Combination therapy of a hif-2-alpha inhibitor and an immunotherapeutic agent and uses thereof

ABSTRACT

The present invention provides methods and pharmaceutical compositions for treating proliferative disorders. The method involves step of administering to said subject a HIF-2alpha inhibitor and an immunotherapeutic agent.

CROSS REFERENCE TO RELATED APPLICATION

This application claims the benefit of U.S. Provisional Application No.62/149,453, filed on Apr. 17, 2015, and U.S. Provisional Application No.62/219,039, filed on Sep. 15, 2015, each incorporated herein byreference in its entirety.

BACKGROUND OF THE INVENTION

An adequate supply of oxygen to tissues is essential in maintainingmammalian cell function and physiology. A deficiency in oxygen supply totissues is a characteristic of a number of pathophysiologic conditionsin which there is insufficient blood flow to provide adequateoxygenation, for example, ischemic disorders, cancer, andatherosclerosis. The hypoxic (low oxygen) environment of tissuesactivates a signaling cascade that drives the induction or repression ofthe transcription of a multitude of genes implicated in events such asangiogenesis (neo-vascularization), glucose metabolism, and cellsurvival/death. A key to this hypoxic transcriptional response lies inthe transcription factors, the hypoxia-inducible factors (HIF). HIFs aredisregulated in a vast array of cancers through hypoxia-dependent andindependent mechanisms and expression is associated with poor patientprognosis.

HIFs consist of an oxygen-sensitive HIFα subunit and a constitutivelyexpressed HIFβ subunit. When HIFs are activated, the HIFα and HIFβsubunits assemble a functional heterodimer (the a subunitheterodimerizes with the β subunit). Both HIFα and HIFβ have twoidentical structural characteristics, a basic helix-loop-helix (bHLH)and PAS domains (PAS is an acronym referring to the first proteins, PER,ARNT, SIM, in which this motif was identified). There are three humanHIFα subunits (HIF-1α, HIF-2α, and HIF-3α) that are oxygen sensitive.Among the three subunits, HIF-1a is the most ubiquitously expressed andinduced by low oxygen concentrations in many cell and tissue types.HIF-2α is highly similar to HIF-1α in both structure and function, butexhibits more restricted cell and tissue-specific expression, and mightalso be differentially regulated by nuclear translocation. HIF-3α alsoexhibits conservation with HIF-1α and HIF-2α in the HLH and PAS domains.HIF-1β (also referred to as ARNT—Aryl Hydrocarbon Receptor NuclearTranslocator), the dimerization partner of the HIFα subunits, isconstitutively expressed in all cell types and is not regulated byoxygen concentration.

PD-1 (Programmed cell death protein-1) is a cell surface co-inhibitoryreceptor expressed mainly on T cells and B cells. PD-1 has two knownligands: PD-L1 and PD-L2. The PD-1/PD-L1 signaling axis plays animportant role in the negative regulation of the immune system toprevent autoimmunity and promote self-tolerance. PD-L1 has been thoughtto be the principle mediator of PD-1 dependent immunosuppression.

PD-L1 is expressed in many human cancers and is associated with a poorprognosis for patients. Tumor-infiltrating lymphocytes from patientswith cancer typically express PD-1 and display impaired antitumorfunctionality. Preclinical studies demonstrate that blockage of theinteraction between PD-1 and PD-L1 can enhance T-cell function andmediate anti-tumor activity. Monoclonal antibodies targeting PD-1 orPD-L1 are under development for the treatment of cancer. One such PD-1targeting agent, nivolumab (Opdivo, Bristol-Myers Squibb), producedcomplete or partial antitumor responses in multiple diseases, includingnon-small-cell lung cancer, melanoma and renal-cell cancer in a clinicaltrial, and was approved by the FDA to treat metastatic melanoma in 2014.There are at least 7 mAbs that target the PD-1/PD-L1 interaction beingevaluated in clinical studies that hold promise for theimmunotherapeutic treatment of various malignancies.

Preclinical and early clinical studies demonstrated that the combinationof anti-PD-1/PD-L1 agents with other targeted agents could increaseanti-tumor efficacy. Simultaneous treatment with anti-PD-1 andanti-VEGFR2 mAbs inhibited tumor growth synergistically in a preclinicalmurine colon cancer model. The most recent phase I clinical trial datashowed that combining nivolumab with one of the current standard of careVEGFR tyrosine kinase inhibitors (TKIs), either sunitinib or pazopanib,significantly increased the overall response rate compared with thesingle agent alone in patients with renal cancer.

CTLA-4 (Cytotoxic T-lymphocyte-associated antigen 4) is a member of theimmunoglobulin superfamily, which is expressed on the surface of Tcells. CTLA-4 transmits an inhibitory signal to T cells by outcompetingthe T-cell co-stimulatory molecule CD28 for B7 ligands (CD80 and CD86)on the surface of antigen-presenting cells with higher affinity andavidity. In preclinical studies, blockade of CTLA-4 led to a significantin T-cell proliferation and interleukin-2 production.

In addition to the suppression of the effector T cells, CTLA-4 alsoincreases the function of immunosuppressive T regulatory T cells(Tregs). CTLA-4 deficiency in Tregs was shown to diminish thesuppressive capacity of Tregs, and CTLA-4 blockade has been shown todeplete intratumoral Tregs in preclinical models.

Based on these preclinical findings, two antibodies that block CTLA-4 inhumans, ipilimumab and tremelimumab, have been tested in the clinic andhave demonstrated significant durable responses in a broad of spectrumof malignancies. The FDA has approved both ipilimumab and tremelimumabfor the treatment of melanoma.

SUMMARY OF THE INVENTION

In view of the foregoing, there exists a need for improved methods fortreating proliferative disorders. This disclosure provides compounds,compositions, and methods that address this need, and provide otheradvantages as well.

In one aspect, the present disclosure provides a method of treating aproliferative disorder in a subject in need thereof. In one embodiment,the method comprises administering to said subject a HIF-2α inhibitorand an immunotherapeutic agent. In some embodiments, the HIF-2αinhibitor and the immunotherapeutic agent yield a synergistic effect intreating the proliferative disorder. In some embodiments, theproliferative disorder is a cancer condition. In some embodiments, thecancer condition is selected from the group consisting of non-small-celllung carcinoma, melanoma, renal cell carcinoma, colorectal cancer,castration-resistant prostate cancer, hepatocellular carcinoma, squamouscell carcinoma of the head and neck, carcinomas of the esophagus, ovary,gastrointestinal tract and breast, and a hematologic malignancy. In someembodiments, the cancer condition is renal cell carcinoma. In someembodiments, the HIF-2α inhibitor and the immunotherapeutic agenttogether are effective in one or more of inhibiting proliferation ofcancer cells, inhibiting metastasis of cancer cells, killing cancercells, modulating an immune response, and reducing severity or incidenceof symptoms associated with the presence of cancer cells. In someembodiments, the HIF-2α inhibitor and the immunotherapeutic agent areadministered sequentially. In some embodiments, the HIF-2α inhibitor andthe immunotherapeutic agent are administered simultaneously. In someembodiments, the HIF-2α inhibitor and the immunotherapeutic agent formpart of the same composition. In some embodiments, the HIF-2α inhibitorand the immunotherapeutic agent are provided in one or more unit doses.In some embodiments, the HIF-2α inhibitor or the immunotherapeutic agentare administered parenterally, orally, intraperitoneally, intravenously,intraarterially, transdermally, intramuscularly, liposomally, via localdelivery by catheter or stent, subcutaneously, intraadiposally, orintrathecally. In some embodiments, the HIF-2α inhibitor inhibits one ormore biological effects selected from the group consisting ofheterodimerization of HIF-2α to HIF-113, HIF-2α target gene expression,VEGF gene expression, and VEGF protein secretion. In some embodiments,the HIF-2α inhibitor inhibits heterodimerization of HIF-2α to HIF-1β butnot heterodimerization of HIF-1α to HIF-1β. In some embodiments, theHIF-2α inhibitor binds the PAS-B domain cavity of HIF-2α.

In one aspect, the present invention provides a compound of Formula I:

or a pharmaceutically acceptable salt or prodrug thereof, wherein:

X is CR⁵ or N;

Y is CR⁶ or N;

Z is —O—, —S—, —C(HR⁷)—, —N(R⁸)—, or absent;

R¹ is alkyl, heteroalkyl, cycloalkyl, heterocycloalkyl, aryl,heteroaryl, acyl or cyano;

R² is nitro, carboxaldehyde, carboxyl, ester, amido, cyano, halo,sulfonyl, alkyl, alkenyl, alkynyl or heteroalkyl;

R³ is hydrogen, halo, cyano, alkyl, heteroalkyl, alkenyl, alkynyl,amino, carboxaldehyde, carboxylic acid, oxime, ester, amido or acyl; orR² and R³ taken together form a cyclic moiety;

R⁴ is nitro, halo, cyano, alkyl, cycloalkyl, heteroaryl, carboxyl,sulfinyl, sulfonamidyl, sulfonyl or sulfoximinyl; and

R⁵, R⁶, R⁷ and R⁸ are independently hydrogen, halo, hydroxy, cyano,alkyl or alkoxy.

In some other embodiments of compounds of Formula I, R¹ is phenyl,monocyclic heteroaryl or bicyclic heteroaryl. In some embodiments, R¹ isphenyl or pyridyl. In yet other embodiments, R¹ is cycloalkyl orheterocycloalkyl. Compounds of Formula I are also provided wherein R¹ issubstituted with at least one substituent selected from the groupconsisting of halo, C₁-C₄ alkyl, C₁-C₄ alkoxy and cyano.

In yet other embodiments of compounds of Formula I, R² and R³ areindependently selected from halo, cyano and alkyl. In some furtherembodiments, R³ is —(CH₂)_(n)OH and n is 1, 2 or 3. In still otherembodiments, n is 1.

In still other embodiments of compounds of Formula I, R⁴ is cyano,fluoroalkyl, sulfinyl, sulfonamidyl, sulfonyl or sulfoximinyl. In otherembodiments, R⁴ is fluoroalkyl or alkylsulfonyl.

In some embodiments of compounds of Formula I, R² and R³ areindependently selected from halo, cyano and alkyl; and R⁴ is cyano,fluoroalkyl, sulfinyl, sulfonamidyl, sulfonyl or sulfoximinyl. Inanother embodiment, R³ is CH₂OH.

In yet another embodiment, Z is —O—. In some embodiments, Z is —S—. Instill other embodiments, Z is —N(R⁸)—. In another embodiment, Z is—C(HR⁷)—. In some embodiments, Z is absent. In some embodiments, X is Nand Y is CR⁶. In another embodiment, X is CR⁵ and Y is N. In yet anotherembodiment, X is N and Y is N. In still another embodiment, X is CR⁵ andY is CR⁶.

In another aspect, the invention provides a compound of Formula I-A:

or a pharmaceutically acceptable salt or prodrug thereof, wherein:

X is CR⁵ or N;

Y is CR⁶ or N;

Z is —O—, —S—, —C(HR⁷)—, —N(R⁸)—, or absent;

R² is nitro, carboxaldehyde, carboxylic acid, ester, amido, cyano, halo,sulfonyl or alkyl;

R³ is hydrogen, halo, cyano, alkyl, heteroalkyl, alkenyl, alkynyl,amino, oxime or acyl; or R² and R³ taken together form a cyclic moiety;

R⁴ is nitro, halo, cyano, alkyl, sulfinyl, sulfonamidyl, sulfonyl orsulfoximinyl;

R⁵, R⁶, R⁷ and R⁸ are independently hydrogen, halo, hydroxy, cyano,alkyl or alkoxy;

W¹ is N or CR¹⁰;

R⁹ is cyano, halo, alkyl or alkoxy; and

R¹⁰ is hydrogen, cyano, halo, alkyl or alkoxy.

In another aspect, the invention provides a compound of Formula I-B:

or a pharmaceutically acceptable salt or prodrug thereof, wherein:

X is CR⁵ or N;

Y is CR⁶ or N;

Z is —O—, —S—, —C(HR⁷)—, —N(R⁸)—, or absent;

R² is nitro, carboxaldehyde, carboxylic acid, ester, amido, cyano, halo,sulfonyl or alkyl;

R³ is hydrogen, halo, cyano, alkyl, heteroalkyl, alkenyl, alkynyl,amino, oxime or acyl; or R² and R³ taken together form a cyclic moiety;

R⁴ is nitro, halo, cyano, alkyl, sulfinyl, sulfonamidyl, sulfonyl orsulfoximinyl;

R⁵, R⁶, R⁷ and R⁸ are independently hydrogen, halo, hydroxy, cyano,alkyl or alkoxy;

R^(c) is hydrogen, cyano, halo, alkyl or alkoxy; and

n′ is 0, 1, 2, 3 or 4.

In yet another aspect, the invention provides a compound of Formula I-C:

or a pharmaceutically acceptable salt or prodrug thereof, wherein:

X is CR⁵ or N;

Y is CR⁶ or N;

Z is —O—, —S—, —C(HR⁷)—, —N(R⁸)—, or absent;

R¹ is alkyl, heteroalkyl, cycloalkyl, heterocycloalkyl, aryl,heteroaryl, acyl or cyano;

R⁴ is nitro, halo, cyano, alkyl, cycloalkyl, heteroaryl, carboxyl,sulfinyl, sulfonamidyl, sulfonyl or sulfoximinyl;

R⁵, R⁶, R⁷ and R⁸ are independently hydrogen, halo, hydroxy, cyano,alkyl or alkoxy;

R¹¹ is hydrogen, hydroxy, alkoxy or amino;

R¹² is hydrogen, alkyl, alkenyl or alkynyl; or R¹¹ and R¹² incombination form oxo or oxime;

each of R¹³ is independently selected from the group consisting ofhydrogen, fluoro, chloro, hydroxy, alkyl and heteroalkyl, with theproviso that when R¹³ is hydroxy, n is 1 or 2; or two R¹³s and thecarbon atom(s) to which they are attached form a 3- to 8-memberedcycloalkyl or heterocycloalkyl moiety; and

n is 0, 1, 2, 3 or 4.

In some other embodiments of compounds of Formula I-C, R¹ is phenyl,monocyclic heteroaryl or bicyclic heteroaryl. In some embodiments, R¹ isphenyl or pyridyl. In yet other embodiments, R¹ is cycloalkyl orheterocycloalkyl. Compounds of Formula I-C are also provided wherein R¹is substituted with at least one substituent selected from the groupconsisting of halo, C₁-C₄ alkyl, C₁-C₄ alkoxy and cyano.

In still other embodiments of compounds of Formula I-C, R⁴ is cyano,fluoroalkyl, sulfinyl, sulfonamidyl, sulfonyl or sulfoximinyl. In otherembodiments, R⁴ is fluoroalkyl or alkylsulfonyl. In some otherembodiments, R¹¹ is hydroxy or amino. In further embodiments, R¹¹ ishydroxy. In yet other embodiments, R¹² is hydrogen. In yet anotherembodiment, R¹³ is fluoro and n is 1, 2 or 3.

In some embodiments of compounds of Formula I-C, R⁴ is cyano,fluoroalkyl, sulfinyl, sulfonamidyl, sulfonyl or sulfoximinyl; R¹¹ ishydroxy or amino; and R¹² is hydrogen. In still another embodiment, R¹³is fluoro. In yet another embodiment, Z is —O—. In some embodiments, Zis —S—. In still other embodiments, Z is —N(R⁸)—. In another embodiment,Z is —C(HR⁷). In some embodiments, Z is absent.

In some other embodiments of compounds of Formula I-C, R⁴ isfluoroalkyl; n is 0, 1, 2 or 3; Z is —O—; R¹¹ is hydroxy; and R¹² ishydrogen. In still other embodiments, R⁴ is sulfonyl; n is 0, 1, 2 or 3;Z is —O—; R¹¹ is hydroxy; and R¹² is hydrogen. In yet other embodiments,R¹ is phenyl, pyridyl, cycloalkyl or heterocycloalkyl. In someembodiments, X is N and Y is CR⁶. In another embodiment, X is CR⁵ and Yis N. In yet another embodiment, X is N and Y is N.

In still another aspect, the invention provides a compound of FormulaI-D, I-E, I-F or I-G:

or a pharmaceutically acceptable salt or prodrug thereof, wherein:

X is CR⁵ or N;

Y is CR⁶ or N;

Z is —O—, —S—, —C(HR⁷)—, —N(R⁸)—, or absent;

R¹ is alkyl, heteroalkyl, cycloalkyl, heterocycloalkyl, aryl orheteroaryl;

R⁴ is nitro, halo, cyano, alkyl, cycloalkyl, heteroaryl, carboxyl,sulfinyl, sulfonamidyl, sulfonyl or sulfoximinyl;

R⁵, R⁶, R⁷ and R⁸ are independently hydrogen, halo, hydroxy, cyano,alkyl or alkoxy; and

R¹¹ is hydrogen, hydroxy, alkoxy or amino.

In a further aspect, the invention provides a compound of Formula I-H,I-I, I-J or I-K:

or a pharmaceutically acceptable salt or prodrug thereof, wherein:

X is CR⁵ or N;

Y is CR⁶ or N;

Z is —O—, —S—, —C(HR⁷)—, —N(R⁸)—, or absent;

R¹ is alkyl, heteroalkyl, cycloalkyl, heterocycloalkyl, aryl orheteroaryl;

R⁴ is nitro, halo, cyano, alkyl, cycloalkyl, heteroaryl, carboxyl,sulfinyl, sulfonamidyl, sulfonyl or sulfoximinyl;

R⁵, R⁶, R⁷ and R⁸ are independently hydrogen, halo, hydroxy, cyano,alkyl or alkoxy; and

R¹¹ is hydroxy or amino.

In yet another aspect, the invention provides a compound of Formula II:

or a pharmaceutically acceptable salt or prodrug thereof, wherein:

Z is —O—, —S—, —C(HR⁷)—, —N(R⁸)—, or absent;

R¹ is alkyl, heteroalkyl, cycloalkyl, heterocycloalkyl, aryl,heteroaryl, acyl or cyano;

R² is hydrogen, alkyl, heteroalkyl, alkenyl, alkynyl, nitro,carboxaldehyde, carboxylic acid, ester, amido, cyano, halo or sulfonyl;

R¹⁴ is hydrogen, deuterium or alkyl;

R¹⁵ is hydrogen, hydroxy or amino; or R¹⁴ and R¹⁵ in combination formoxo or methylene;

R¹⁶ and R¹⁷ are independently selected from the group consisting ofhydrogen, halo, alkyl, heteroalkyl and cycloalkyl; or R¹⁶ and R¹⁷ andthe carbon to which they are attached form C₃-C₈ cycloalkyl or C₅-C₈heterocycloalkyl;

R¹⁸ is O or NR¹⁹, wherein R¹⁹ is selected from the group consisting ofhydrogen, alkyl and cyano;

n″ is 1 or 2; and

R⁷ and R⁸ are independently hydrogen, halo, hydroxy, cyano, alkyl oralkoxy.

In another aspect, the invention provides a compound of Formula II-A:

or a pharmaceutically acceptable salt or prodrug thereof, wherein:

Z is —O—, —S—, —C(HR⁷)—, —N(R⁸)—, or absent;

R¹ is alkyl, heteroalkyl, cycloalkyl, heterocycloalkyl, aryl,heteroaryl, acyl or cyano;

R² is hydrogen, alkyl, heteroalkyl, alkenyl, alkynyl, nitro,carboxaldehyde, carboxylic acid, ester, amido, cyano, halo or sulfonyl;

R¹⁴ is hydrogen, deuterium or alkyl;

R¹⁵ is hydrogen, hydroxy or amino; or R¹⁴ and R¹⁵ in combination formoxo or methylene;

R¹⁸ is O or NR¹⁹, wherein R¹⁹ is selected from the group consisting ofhydrogen, alkyl and cyano; and

R⁷ and R⁸ are independently hydrogen, halo, hydroxy, cyano, alkyl oralkoxy.

In still another aspect, the invention provides a compound of FormulaII-B:

or a pharmaceutically acceptable salt or prodrug thereof, wherein:

Z is —O—, —S—, —C(HR⁷)—, —N(R⁸)—, or absent;

R¹ is alkyl, heteroalkyl, cycloalkyl, heterocycloalkyl, aryl,heteroaryl, acyl or cyano;

R² is hydrogen, alkyl, heteroalkyl, alkenyl, alkynyl, nitro,carboxaldehyde, carboxylic acid, ester, amido, cyano, halo or sulfonyl;

R¹⁵ is hydroxy or amino;

R¹⁸ is O or NR¹⁹, wherein R¹⁹ is selected from the group consisting ofhydrogen, alkyl and cyano; and

R⁷ and R⁸ are independently hydrogen, halo, hydroxy, cyano, alkyl oralkoxy.

In some embodiments, the present invention provides a HIF-2α inhibitorthat may be a compound selected from Table 1. In some embodiments, theimmunotherapeutic agent induces or enhances an immune response. In someembodiments, the immunotherapeutic agent is a PD-1 inhibitor. The PD-1inhibitor may be nivolumab or pembrolizumab. In some embodiments, theimmunotherapeutic agent is a CTLA-4 inhibitor. The CTLA-4 inhibitor maybe tremelimumab or ipilimumab. In some embodiments, theimmunotherapeutic agent comprises a small molecule or antibody. In someembodiments, the immunotherapeutic agent comprises an antibody. In someembodiments, the present invention provides a method of measuring atumor size or an amount of one or more markers for the presence of theproliferative disorder before and after administering the HIF-2αinhibitor and the immunotherapeutic agent, and (b) adjusting dosage ordiscontinuing use of the HIF-2α inhibitor and/or the immunotherapeuticagent based on results of step (a).

INCORPORATION BY REFERENCE

All publications and patent applications mentioned in this specificationare herein incorporated by reference to the same extent as if eachindividual publication or patent application was specifically andindividually indicated to be incorporated by reference.

BRIEF DESCRIPTION OF THE DRAWINGS

FIG. 1 shows a synergistic effect of a treatment according to themethods of the invention on tumor growth in mice.

FIG. 2 depicts results of a co-immunoprecipitation assay for measuringinhibition of HIF-2α and ARNT dimerization.

FIG. 3 shows a synergistic effect of a treatment according to themethods of the invention on tumor growth in mice.

DETAILED DESCRIPTION

While preferred embodiments of the present invention have been shown anddescribed herein, it will be obvious to those skilled in the art thatsuch embodiments are provided by way of example only. Numerousvariations, changes and substitutions will now occur to those skilled inthe art without departing from the invention. It should be understoodthat various alternatives to the embodiments of the invention describedherein may be employed in practicing the invention. It is intended thatthe appended claims define the scope of the invention and that methodsand structures within the scope of these claims and their equivalents becovered thereby.

Unless defined otherwise, all technical and scientific terms used hereinhave the same meaning as is commonly understood by one of skill in theart to which this invention belongs. All patents and publicationsreferred to herein are incorporated by reference.

As used in the specification and claims, the singular form “a”, “an” and“the” includes plural references unless the context clearly dictatesotherwise.

As used herein, “agent” or “biologically active agent” refers to abiological, pharmaceutical, or chemical compound or other moiety.Non-limiting examples include a simple or complex organic or inorganicmolecule, a peptide, a protein, an oligonucleotide, an antibody, anantibody derivative, antibody fragment, a vitamin derivative, acarbohydrate, a toxin, or a chemotherapeutic compound. Various compoundscan be synthesized, for example, small molecules and oligomers (e.g.,oligopeptides and oligonucleotides), and synthetic organic compoundsbased on various core structures. In addition, various natural sourcescan provide compounds for screening, such as plant or animal extracts,and the like. A skilled artisan can readily recognize that there is nolimit as to the structural nature of the agents of the presentinvention.

The terms “antagonist” and “inhibitor” are used interchangeably, andthey refer to a compound having the ability to inhibit a biologicalfunction of a target protein, whether by inhibiting the activity orexpression of the target protein. Accordingly, the terms “antagonist”and “inhibitors” are defined in the context of the biological role ofthe target protein. While preferred antagonists herein specificallyinteract with (e.g., bind to) the target, compounds that inhibit abiological activity of the target protein by interacting with othermembers of the signal transduction pathway of which the target proteinis a member are also specifically included within this definition. Apreferred biological activity inhibited by an antagonist is associatedwith the development, growth, or spread of a tumor, or an undesiredimmune response as manifested in autoimmune disease.

The term “cell proliferation” refers to a phenomenon by which the cellnumber has changed as a result of division. This term also encompassescell growth by which the cell morphology has changed (e.g., increased insize) consistent with a proliferative signal.

The terms “co-administration,” “administered in combination with,” andtheir grammatical equivalents, encompass administration of two or moreagents to an animal so that both agents and/or their metabolites arepresent in the animal at the same time. Co-administration includessimultaneous administration in separate compositions, administration atdifferent times in separate compositions, or administration in acomposition in which both agents are present.

The term “effective amount” or “therapeutically effective amount” refersto that amount of a compound described herein that is sufficient toeffect the intended application including but not limited to diseasetreatment, as defined below. The therapeutically effective amount mayvary depending upon the intended application (in vitro or in vivo), orthe subject and disease condition being treated, e.g., the weight andage of the subject, the severity of the disease condition, the manner ofadministration and the like, which can readily be determined by one ofordinary skill in the art. The term also applies to a dose that willinduce a particular response in target cells, e.g., reduction ofplatelet adhesion and/or cell migration. The specific dose will varydepending on the particular compounds chosen, the dosing regimen to befollowed, whether it is administered in combination with othercompounds, timing of administration, the tissue to which it isadministered, and the physical delivery system in which it is carried.

As used herein, the terms “treatment”, “treating”, “palliating” and“ameliorating” are used interchangeably. These terms refer to anapproach for obtaining beneficial or desired results including, but arenot limited to, therapeutic benefit and/or a prophylactic benefit. Bytherapeutic benefit is meant eradication or amelioration of theunderlying disorder being treated. Also, a therapeutic benefit isachieved with the eradication or amelioration of one or more of thephysiological symptoms associated with the underlying disorder such thatan improvement is observed in the patient, notwithstanding that thepatient can still be afflicted with the underlying disorder. Forprophylactic benefit, the pharmaceutical compositions may beadministered to a patient at risk of developing a particular disease, orto a patient reporting one or more of the physiological symptoms of adisease, even though a diagnosis of this disease may not have been made.

A “therapeutic effect,” as used herein, encompasses a therapeuticbenefit and/or a prophylactic benefit as described above. A prophylacticeffect includes delaying or eliminating the appearance of a disease orcondition, delaying or eliminating the onset of symptoms of a disease orcondition, slowing, halting, or reversing the progression of a diseaseor condition, or any combination thereof.

The term “pharmaceutically acceptable” means that a chemical entity,such as a compound, a carrier, an additive or a salt, is acceptable forbeing administrated to a subject.

The term “pharmaceutically acceptable salt” refers to salts derived froma variety of organic and inorganic counter ions well known in the art.Pharmaceutically acceptable acid addition salts can be formed withinorganic acids and organic acids. Inorganic acids from which salts canbe derived include, for example, hydrochloric acid, hydrobromic acid,sulfuric acid, nitric acid, phosphoric acid, and the like. Organic acidsfrom which salts can be derived include, for example, acetic acid,propionic acid, glycolic acid, pyruvic acid, oxalic acid, maleic acid,malonic acid, succinic acid, fumaric acid, tartaric acid, citric acid,benzoic acid, cinnamic acid, mandelic acid, methanesulfonic acid,ethanesulfonic acid, p-toluenesulfonic acid, salicylic acid, and thelike. Pharmaceutically acceptable base addition salts can be formed withinorganic and organic bases. Inorganic bases from which salts can bederived include, for example, sodium, potassium, lithium, ammonium,calcium, magnesium, iron, zinc, copper, manganese, aluminum, and thelike. Organic bases from which salts can be derived include, forexample, primary, secondary, and tertiary amines, substituted aminesincluding naturally occurring substituted amines, cyclic amines, basicion exchange resins, and the like, specifically such as isopropylamine,trimethylamine, diethylamine, triethylamine, tripropylamine, andethanolamine. In some embodiments, the pharmaceutically acceptable baseaddition salt is chosen from ammonium, potassium, sodium, calcium, andmagnesium salts.

The term “pharmaceutically acceptable carrier” or “pharmaceuticallyacceptable excipient” includes any and all solvents, dispersion media,coatings, antibacterial and antifungal agents, isotonic and absorptiondelaying agents and the like. The use of such media and agents forpharmaceutically active substances is well known in the art. Exceptinsofar as any conventional media or agent is incompatible with theactive ingredient, its use in the therapeutic compositions of theinvention is contemplated. Supplementary active ingredients can also beincorporated into the compositions.

The term “selective inhibition” or “selectively inhibit” as applied to abiologically active agent refers to the agent's ability to selectivelyreduce the target signaling activity as compared to off-target signalingactivity, via direct or indirect interaction with the target.

The term “subject” includes, but is not limited to, humans of any agegroup, e.g., a pediatric subject (e.g., infant, child or adolescent) oradult subject (e.g., young adult, middle-aged adult or senior adult))and/or other primates (e.g., cynomolgus monkeys or rhesus monkeys);mammals, including commercially relevant mammals such as cattle, pigs,horses, sheep, goats, cats, and/or dogs; and/or birds, includingcommercially relevant birds such as chickens, ducks, geese, quail,and/or turkeys. The methods described herein can be useful in both humantherapeutics and veterinary applications. In some embodiments, thepatient is a mammal, and in some embodiments, the patient is human.

“Radiation therapy” or “radiation treatment” means exposing a patient,using routine methods and compositions known to the practitioner, toradiation emitters such as alpha-particle emitting radionucleotides(e.g., actinium and thorium radionuclides), low linear energy transfer(LET) radiation emitters (e.g., beta emitters), conversion electronemitters (e.g., strontium-89 and samarium-153-EDTMP), or high-energyradiation, including without limitation x-rays, gamma rays, andneutrons.

The term “immunotherapeutic agent” refers to any agent that induces,enhances, suppresses or otherwise modifies an immune response. Thisincludes the administration of an active agent to, or any type ofintervention or process performed on, the subject, with the objective ofmodifying an immune response. An immunotherapeutic agent may, forexample, increase or enhance the effectiveness or potency of an existingimmune response in a subject, for example, by stimulating mechanismsthat enhance the endogenous host immune response or overcomingmechanisms that suppress the endogenous host immune response.

“Immune response” refers to the action of a cell of the immunesystem-including, for example, B lymphocytes, T lymphocytes, naturalkiller (NK) cells, macrophages, eosinophils, mast cells, myeloid-derivedsuppressor cells, dendritic cells and neutrophils- and solublemacromolecules produced by any of these cells or the liver (includingantibodies, cytokines and complement), that results in selectivetargeting, binding to, damage to, destruction of, and/or elimination ofinvading pathogens, cells or tissues infected with pathogens, cancerousor other abnormal cells, or, in cases of autoimmunity or pathologicalinflammation, normal human cells or tissues, from the body of a subject.

The term “antibody” (Ab) as referred to herein includes whole antibodiesand any antigen binding fragment (i.e. “antigen-binding portion”) orsingle chains thereof. A typical full-length antibody is a glycoproteinimmunoglobulin which binds specifically to an antigen and comprises atleast two heavy (H) chains and two light (L) chains interconnected bydisulfide bonds, or an antigen-binding portion thereof. Each H chaincomprises a heavy chain variable region (abbreviated herin as V_(H)) anda heavy chain constant region. The heavy chain constant region comprisesthree constant domains, C_(H1), C_(H2) and C_(H3). Each light chaincomprises a light chain variable region (abbreviated herein as V_(L))and a light chain constant region. The light chain constant regioncomprises one constant domain, C_(L). The V_(H) and V_(L) regions can befurther subdivided into regions of hypervariability, termedcomplementarity determining regions (CDRs), interspersed with regionsthat are more conserved, termed framework regions (FR). Each V_(H) andV_(L) comprises three CDRs and four FRs, arranged from amino-terminus tocarboxy-terminus in the following order: FR1, CDR1, FR2, CDR2, FR3,CDR3, FR4. The variable regions of the heavy and light chains contain abinding domain that interacts with an antigen. The constant regions ofthe Abs may mediate the binding of the immunoglobulin to host tissues orfactors, including various cells of the immune system (e.g., effectorcells) and the first component (Clq) of the classical complement system.

Antibodies typically bind specifically to their cognate antigen withhigh affinity, reflected by a dissociation constant (K_(D)) of 10⁻⁵ to10⁻¹¹ M⁻¹ or less. In some embodiments, an Ab that “binds specifically”to an antigen refers to an Ab that binds to the antigen andsubstantially identical antigens with a K_(D) of 10⁻⁷ M⁻¹ or less,preferably 10⁻⁸ M⁻¹ or less, even more preferably 5×10⁻⁹ M⁻¹ or less,and most preferably between 10⁻⁸ M⁻¹ and 10⁻¹⁰ M⁻¹ or less, but does notbind with such affinity to unrelated antigens. An antigen is“substantially identical” to a given antigen if it exhibits a highdegree of sequence identity to the given antigen, for example, if itexhibits at least 80%, at least 90%, preferably at least 95%, morepreferably at least 97%, or even more preferably at least 99% sequenceidentity to the sequence of the given antigen. By way of example, an Abthat binds specifically to human PD-1 may also have cross-reactivitywith PD-1 antigens from certain primate species but may not cross-reactwith PD-1 antigens from certain rodent species or with an antigen otherthan PD-1, e.g., a human PD-L1 antigen.

An immunoglobulin may derive from any of the commonly known isotypes,including but not limited to IgA, secretory IgA, IgG and IgM. IgGsubclasses are also well known to those in the art and include, but arenot limited to, human IgG1, IgG2, IgG3 and IgG4. “Isotype” refers to theAb class or subclass (e.g., IgM or IgG1) that is encoded by the heavychain constant region genes. The term “antibody” includes, by way ofexample, both naturally occurring and non-naturally occurring Abs;monoclonal and polyclonal Abs; chimeric and humanized Abs; human ornonhuman Abs; wholly synthetic Abs; and single chain Abs. A nonhuman Abmay be humanized by recombinant methods to reduce its immunogenicity inman. Where not expressly stated, and unless the context indicatesotherwise, the term “antibody” also includes an antigen-binding fragmentor an antigen-binding portion of any of the aforementionedimmunoglobulins, and includes a monovalent and a divalent fragment orportion, and a sing chain Ab.

An “isolated antibody” refers to an Ab that is substantially free ofother Abs having different antigenic specificities (e.g., an isolated Abthat binds specifically to PD-1 is substantially free of Abs that bindspecifically to antigens other than PD-1). An isolated Ab that bindsspecifically to PD-1 may, however, have cross-reactivity to otherantigens, such as PD-1 molecules from different species. Moreover, anisolated Ab may be substantially free of other cellular material and/orchemicals.

The term “monoclonal antibody” (“mAb”) refers to a preparation of Abmolecules of single molecular composition, i.e., Ab molecules whoseprimary sequences are essentially identical, and which exhibits a singlebinding specificity and affinity for a particular epitope. A mAb is anexample of an isolated Ab. MAbs may be produced by hybridoma,recombinant, transgenic or other techniques known to those skilled inthe art.

A “human” antibody (HuMAb) refers to an Ab having variable regions inwhich both the framework and CDR regions are derived from human germlineimmunoglobulin sequences. Furthermore, if the Ab contains a constantregion, the constant region also is derived from human germlineimmunoglobulin sequences. The human Abs of the invention may includeamino acid residues not encoded by human germline immunoglobulinsequences (e.g., mutations introduced by random or site-specificmutagenesis in vitro or by somatic mutation in vivo). However, the term“human antibody,” as used herein, is not intended to include Abs inwhich CDR sequences derived from the germline of another mammalianspecies, such as a mouse, have been grafted onto human frameworksequences. The terms “human” Abs and “fully human” Abs and are usedsynonymously.

A “humanized” antibody refers to an Ab in which some, most or all of theamino acids outside the CDR domains of a non-human Ab are replaced withcorresponding amino acids derived from human immunoglobulins. In oneembodiment of a humanized form of an Ab, some, most or all of the aminoacids outside the CDR domains have been replaced with amino acids fromhuman immunoglobulins, whereas some, most or all amino acids within oneor more CDR regions are unchanged. Small additions, deletions,insertions, substitutions or modifications of amino acids arepermissible as long as they do not abrogate the ability of the Ab tobind to a particular antigen. A “humanized” Ab retains an antigenicspecificity similar to that of the original Ab.

A “chimeric antibody” refers to an Ab in which the variable regions arederived from one species and the constant regions are derived fromanother species, such as an Ab in which the variable regions are derivedfrom a mouse Ab and the constant regions are derived from a human Ab.

An “antigen-binding portion” of an Ab (also called an “antigen-bindingfragment”) refers to one or more fragments of an Ab that retain theability to bind specifically to the antigen bound by the whole Ab.

The “Programmed Death-1 (PD-1)” receptor refers to an immunoinhibitoryreceptor belonging to the CD28 family. PD-1 is expressed predominantlyon previously activated T cells in vivo, and binds to two ligands, PD-L1and PD-L2. The term “PD-1” as used herein includes human PD-1 (hPD-1),variants, isoforms, and species homologs of hPD-1, and analogs having atleast one common epitope with hPD-1. The complete hPD-1 sequence can befound under GENBANK® Accession No. U64863.

“Programmed Death Ligand-1 (PD-L1)” is one of two cell surfaceglycoprotein ligands for PD-1 (the other being PD-L2) that downregulateT cell activation and cytokine secretion upon binding to PD-1. The term“PD-L1” as used herein includes human PD-L1 (hPD-L1), variants,isoforms, and species homologs of hPD-L1, and analogs having at leastone common epitope with hPD-L1. The complete hPD-L1 sequence can befound under GENBANK® Accession No. Q9NZQ7.

The terms “cytotoxic T lymphocyte-associated antigen-4,” “CTLA-4,”“CTLA4” and “CD152” are used interchangeably, and refer to animmunoinhibitory receptor belonging to the immunoglobulin (Ig)superfamily. CTLA-4 is a cell surface receptor expressed predominantlyon activated T cells, and binds to two ligands, CD80 (B7-1) and CD86(B7-2). The term “CTLA-4” as used herein includes variants, isoforms,species homologs of human CTLA-4, and analogs having at least one commonepitope with CTLA-4. The complete sequence of human CTLA-4 can be foundunder GENBANK® Accession No. L15006.

A “cell surface receptor” includes, for example, molecules and complexesof molecules that are located on the surface of a cell and are capableof receiving a signal and transmitting such a signal across the plasmamembrane of a cell. An example of a cell surface receptor of the presentinvention is the PD-1 receptor, which is located on the surface ofactivated B cells, activated T cells and myeloid cells, and transmits asignal that results in a decrease in tumor-infiltrating lymphocytes anda decrease in T cell proliferation.

A “PD-1 inhibitor” as referred to herein is meant to include any agent,including an antibody or small molecule, that blocks or inhibits thePD-1 pathway. A PD-1 inhibitor may, for example, block or inhibit aninteraction between PD-1/PD-L1, PD-1/PD-L2 and/or PD-L1/CD80. A PD-1inhibitor includes any compound able to directly or indirectly affectthe regulation of PD-1 by reducing, for example, the expression of PD-1,PD-L1 or PD-L2, or a PD-1 activity.

A “CTLA-4 inhibitor” as referred to herein is meant to include anyagent, including an antibody or small molecule, that blocks or inhibitsthe CTLA-4 pathway. A CTLA-4 inhibitor may, for example, block orinhibit an interaction between CTLA-4/CD80 and/or CTLA-4/CD86. A CTLA-4inhibitor includes any compound able to directly or indirectly affectthe regulation of CTLA-4 by reducing, for example, the expression ofCTLA-4, CD80 or CD86, or a CTLA-4 activity.

A “B7 inhibitor” as referred to herein is meant to include any agent,including an antibody or small molecule, that blocks or inhibits aninteraction between a B7 protein and a receptor. “B7 protein” includesmembers of the B7 family, including CD80 (B7-1), CD86 (B7-2), PD-L1(B7-H1) and PD-L2 (B7-DC). Receptors of B7 proteins include PD-1, CTLA-4and CD28. A B7 inhibitor may, for example, block or inhibit aninteraction between PD-1/PD-L1, PD-1/PD-L2, PD-L1/CD80, CTLA-4/CD80and/or CTLA-4/CD86.

“Prodrug” is meant to indicate a compound that may be converted underphysiological conditions or by solvolysis to a biologically activecompound described herein. Thus, the term “prodrug” refers to aprecursor of a biologically active compound that is pharmaceuticallyacceptable. A prodrug may be inactive when administered to a subject,but is converted in vivo to an active compound, for example, byhydrolysis. The prodrug compound often offers advantages of solubility,tissue compatibility or delayed release in a mammalian organism (see,e.g., Bundgard, H., Design of Prodrugs (1985), pp. 7-9, 21-24 (Elsevier,Amsterdam). A discussion of prodrugs is provided in Higuchi, T., et al.,“Pro-drugs as Novel Delivery Systems,” A.C.S. Symposium Series, Vol. 14,and in Bioreversible Carriers in Drug Design, ed. Edward B. Roche,American Pharmaceutical Association and Pergamon Press, 1987, both ofwhich are incorporated in full by reference herein. The term “prodrug”is also meant to include any covalently bonded carriers, which releasethe active compound in vivo when such prodrug is administered to amammalian subject. Prodrugs of an active compound, as described herein,may be prepared by modifying functional groups present in the activecompound in such a way that the modifications are cleaved, either inroutine manipulation or in vivo, to the parent active compound. Prodrugsinclude compounds wherein a hydroxy, amino or mercapto group is bondedto any group that, when the prodrug of the active compound isadministered to a mammalian subject, cleaves to form a free hydroxy,free amino or free mercapto group, respectively. Examples of prodrugsinclude, but are not limited to, acetate, formate and benzoatederivatives of an alcohol or acetamide, formamide and benzamidederivatives of an amine functional group in the active compound and thelike.

The term “in vivo” refers to an event that takes place in a subject'sbody.

The term “in vitro” refers to an event that takes place outside of asubject's body. For example, an in vitro assay encompasses any assay runoutside of a subject's body. In vitro assays encompass cell-based assaysin which cells alive or dead are employed. In vitro assays alsoencompass a cell-free assay in which no intact cells are employed.

The term “HIF-2α” refers to a monomeric protein that contains severalconserved structured domains: basic helix-loop-helix (bHLH), and twoPer-ARNT-Sim (PAS) domains designated PAS-A and PAS-B, in addition toC-terminal regulatory regions. “HIF-2α” is also alternatively known byseveral other names in the scientific literature, including EndothelialPAS Domain Protein 1 (EPAS 1), HIF2A, PASD2, HIF-2-Alpha, HIF2-Alpha,HLF, Hypoxia-Inducible Factor 2-Alpha, HIF-1alpha-Like Factor, and MOP2.As a member of the bHLH/PAS family of transcription factors, “HIF-2α”forms an active heterodimeric transcription factor complex by binding tothe ARNT (also known as HIF-1β) protein through non-covalentinteractions.

The term “scintillation proximity assay” (SPA) refers to a homogeneousassay in which light is emitted when a radiolabeled ligand is broughtinto close proximity to a radiosensitive bead. The assay typicallycontains a target protein that contains a tag (e.g., His Tag,Glutathione S-transferase Tag). The tag on the protein is used to bindthe target protein to the scintillation bead. Radio-labeled ligand(e.g., labeled with tritium) that binds to the protein is now in closeproximity to the bead, and when the radio-label (e.g., tritium) decays,the high energy particle hits the bead resulting in the emission oflight that is detected by a detector, such as photomultiplier tube orCCD camera. When unlabeled ligands or compounds that bind to the proteinare used in the assay, they displace the radio-labeled ligand, resultingin loss of signal. For a general reference describing the assay, seePark, et al. Analytical Biochemistry 269: 94-104, 1999.

“HIF-2α activity” as used herein has its ordinary meaning in the art.HIF-2α activity, for example, includes activation of gene transcriptionmediated by HIF-2α.

The term “inhibiting HIF-2α activity”, as used herein, refers toslowing, reducing, altering, as well as completely eliminating and/orpreventing HIF-2α activity.

“Combination therapy” and “co-therapy” means the administration of afirst active agent and at least a second, different active agent as partof a specific treatment regimen intended to provide the beneficialeffect from the co-action of the at least two active agents. Thebeneficial effect of the combination may include, but is not limited to,pharmacokinetic or pharmacodynamic co-action resulting from thecombination of therapeutic agents. Administration of therapeutic agentsin combination typically is carried out over a defined time period(usually minutes, hours, days or weeks depending upon the combinationselected). Combination therapy is not intended to encompass theadministration of two or more different therapeutic agents as part ofseparate monotherapy regimens that incidentally and arbitrarily resultsin a combination therapy of the invention. Combination therapy includesadministration of at least two different therapeutic agents in asequential manner, wherein each therapeutic agent is administered at adifferent time, as well as administration of at least two differenttherapeutic agents in a substantially simultaneous manner. Substantiallysimultaneous administration can be accomplished, for example, byadministering to the subject a single capsule having a fixed ratio ofeach therapeutic agent or in separate capsules for each of thetherapeutic agents. Sequential or substantially simultaneousadministration of each therapeutic agent can be effected by anyappropriate route, including, but not limited to, oral routes,intravenous routes, intramuscular routes, and direct absorption throughmucous membrane tissues. The two different therapeutic agents can beadministered by the same route or by different routes. For example, afirst therapeutic agent of the combination selected may be administeredby intravenous injection while the second therapeutic agent of thecombination may be administered orally. Alternatively, for example, alltherapeutic agents may be administered orally or all therapeutic agentsmay be administered by intravenous injection. The sequence in which thetherapeutic agents are administered is not critical, unless otherwisestated. Combination therapy also includes the administration of thedifferent therapeutic agents as described above in further combinationwith other biologically active ingredients and non-drug therapies (e.g.,surgery or physical therapy). Where a combination therapy comprises anon-drug treatment, the non-drug treatment may be conducted at anysuitable time so long as a beneficial effect from the co-action of thecombination of the therapeutic agents and non-drug treatment isachieved. For example, in appropriate cases, the beneficial effect isstill achieved when the non-drug treatment is temporally removed fromthe administration of the therapeutic agents, perhaps by days or evenweeks.

The term “simultaneous” or “simultaneously” as applied to administeringmore than one pharmaceutically active ingredient (e.g., a HIF-2αinhibitor and an immunotherapeutic agent) refers to administering themore than one ingredient at the same time, or at two different timepoints that are separated by no more than 2 hours. The term“sequentially” as applied to administering more than onepharmaceutically active ingredient (e.g., a HIF-2α inhibitor and animmunotherapeutic agent) refers to administering the more than oneingredient at two different time points that are separated by more than2 hours, e.g., about 5 hours, 8 hours, 1 day, 2 days, 3 days, 4 days, 5days, 6 days, 7 days or even longer.

As used herein, the term “neoplastic condition” refers to the presenceof cells possessing abnormal growth characteristics, such asuncontrolled proliferation, immortality, metastatic potential, rapidgrowth and proliferation rate, perturbed oncogenic signaling, or certaincharacteristic morphological features.

A “sub-therapeutic amount” of an agent or therapy is an amount less thanthe effective amount for that agent or therapy, but when combined withan effective or sub-therapeutic amount of another agent or therapy canproduce a result desired by the physician, due to, for example, synergyin the resulting efficacious effects, or reduced side effects.

A “synergistic” or “synergizing” effect can be such that the one or moreeffects of the combination compositions are greater than the one or moreeffects of each component alone at a comparable dosing level, or theycan be greater than the predicted sum of the effects of all of thecomponents at a comparable dosing level, assuming that each componentacts independently. The synergistic effect can be about, or greater thanabout 1, 2, 3, 5, 10, 20, 30, 50, 75, 100, 110, 120, 150, 200, 250, 350,or 500% or even more than the effect on a subject with one of thecomponents alone, or the additive effects as measured when each of thecomponents when administered individually. The effect can be any of themeasurable effects described herein.

An “anti-cancer agent”, “anti-tumor agent” or “chemotherapeutic agent”refers to any agent useful in the treatment of a neoplastic condition.One class of anti-cancer agents comprises chemotherapeutic agents.“Chemotherapy” means the administration of one or more chemotherapeuticdrugs and/or other agents to a cancer patient by various methods,including intravenous, oral, intramuscular, intraperitoneal,intravesical, subcutaneous, transdermal, buccal, inhalation, or in theform of a suppository.

“Signal transduction” is a process during which stimulatory orinhibitory signals are transmitted into and within a cell to elicit anintracellular response. A modulator of a signal transduction pathwayrefers to a compound that modulates the activity of one or more cellularproteins mapped to the same specific signal transduction pathway. Amodulator may augment (agonist) or suppress (antagonist) the activity ofa signaling molecule.

The term “heterodimerization” as used herein refers to the complexformed by the non-covalent binding of HIF-2α to HIF-1β (ARNT).Heterodimerization of HIF-2α to HIF-1β (ARNT) is required for HIF-2α DNAbinding and transcriptional activity and is mediated by the HLH andPAS-B domains. Transcriptional activity following heterodimerization ofHIF-2α to HIF-1β(ARNT) can affect four groups of target genes includingangiogenic factors, glucose transporters and glycolytic enzymes,survival factors, and invasion factors.

The term “HIF-2α PAS-B domain cavity” refers to an internal cavitywithin the PAS-B domain of HIF2α. The crystal structure of the PAS-Bdomain can contain a large (approximately 290 A) cavity in its core.However, the amino acid side chains in the solution structure aredynamic. For example, those side chains can tend to intrude more deeplyin the core, and can shrink the cavity to 1 or 2 smaller cavities or caneven expand the cavity. The cavity is lined by amino acid residuescomprising PHE-244, SER-246, HIS-248, MET-252, PHE-254, ALA-277,PHE-280, TYR-281, MET-289, SER-292, HIS-293, LEU-296, VAL-302, VAL-303,SER-304, TYR-307, MET-309, LEU-319, THR-321, GLN-322, GLY-323, ILE-337,CYS-339, and ASN-341 of HIF-2α PAS-B domain. The numbering system isfrom the known structures reported in the RCSB Protein Data Bank withPDB code 3H7W. Other numbering systems in the PDB could define the sameamino acids, expressed above, that line the cavity.

As described herein “a biological marker”, or “a biomarker”, generallyrefers to a measurable indicator of some biological state or condition.Biological markers are often measured and evaluated to examine normalbiological processes, pathogenic processes, or pharmacologic responsesto a therapeutic intervention.

The terms “polynucleotide”, “nucleotide”, “nucleotide sequence”,“nucleic acid” and “oligonucleotide” are used interchangeably. Theyrefer to a polymeric form of nucleotides of any length, eitherdeoxyribonucleotides or ribonucleotides, or analogs thereof.Polynucleotides may have any three dimensional structure, and mayperform any function, known or unknown. The following are non-limitingexamples of polynucleotides: coding or non-coding regions of a gene orgene fragment, loci (locus) defined from linkage analysis, exons,introns, messenger RNA (mRNA), transfer RNA, ribosomal RNA, shortinterfering RNA (siRNA), short-hairpin RNA (shRNA), micro-RNA (miRNA),ribozymes, cDNA, recombinant polynucleotides, branched polynucleotides,plasmids, vectors, isolated DNA of any sequence, isolated RNA of anysequence, nucleic acid probes, and primers. A polynucleotide maycomprise one or more modified nucleotides, such as methylatednucleotides and nucleotide analogs. If present, modifications to thenucleotide structure may be imparted before or after assembly of thepolymer. The sequence of nucleotides may be interrupted bynon-nucleotide components. A polynucleotide may be further modifiedafter polymerization, such as by conjugation with a labeling component.

“Controlled release” refers to a drug-containing formulation or unitdose form thereof from which release of the drug is not immediate, i.e.,with a controlled release formulation, administration does not result inimmediate release of all of the drug administered into an absorptionpool. The term is used interchangeably with “nonimmediate release” asdefined in Remington: The Science and Practice of Pharmacy, NineteenthEd. (Easton, Pa.: Mack Publishing Company, 1995). In general, controlledrelease formulations include sustained release and delayed releaseformulations.

“Sustained release” and “extended release” refers to a drug formulationthat provides for gradual release of a drug over an extended period oftime, and typically, although not necessarily, results in substantiallyconstant blood levels of a drug over an extended time period.

“Delayed release” refers to a drug formulation that, followingadministration to a patient, provides a measurable time delay beforedrug is released from the formulation into the patient's body.

“Dosage form” means any form of a pharmaceutical composition foradministration to a subject (typically a human or animal of veterinaryinterest suffering from a disease or condition to be treated). “Dose”refers to an amount of active agent. “Unit dosage form” refers to adosage form that contains a fixed amount of active agent. A singletablet or capsule is a unit dosage form. Multiple unit dosage forms canbe administered to provide a therapeutically effective dose. A dosageform can include a combination of dosage forms.

The term “alkyl” refers to a straight or branched hydrocarbon chainradical comprising carbon and hydrogen atoms, containing nounsaturation, and having from one to ten carbon atoms (e.g., C₁-C₁₀alkyl). Whenever it appears herein, a numerical range such as “1 to 10”refers to each integer in the given range; e.g., “1 to 10 carbon atoms”means that the alkyl group may consist of 1 carbon atom, 2 carbon atoms,3 carbon atoms, etc., up to and including 10 carbon atoms, although thepresent definition also covers the occurrence of the term “alkyl” whereno numerical range is designated. In some embodiments, it is a C₁-C₄alkyl group. Typical alkyl groups include, but are in no way limited to,methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, sec-butyl,tert-butyl, pentyl, isopentyl, neopentyl, hexyl, septyl, octyl, nonyl,decyl, and the like. The alkyl is attached to the rest of the moleculeby a single bond, for example, methyl (Me), ethyl (Et), n-propyl,1-methylethyl, (iso-propyl), n-butyl, n-pentyl, 1,1-dimethylethyl(t-butyl), 3-methylhexyl, 2-methylhexyl, and the like. Unless statedotherwise specifically in the specification, an alkyl group isoptionally substituted by one or more of substituents whichindependently are: alkyl, heteroalkyl, alkenyl, alkynyl, cycloalkyl,heterocycloalkyl, aryl, arylalkyl, heteroaryl, heteroarylalkyl, hydroxy,halo, cyano, trifluoromethyl, trifluoromethoxy, nitro, oxo, thioxo,trimethylsilanyl, —OR^(a), —SR^(a), —OC(═O)R^(a), —OC(═O)OR^(a),—OC(═O)N(R^(a))₂, —N(R^(a))₂, —C(═O)OR^(a), —C(═O)R^(a),—C(═O)N(R^(a))₂, —N(R^(a))C(═O)OR^(a), —N(R^(a))C(═O)N(R^(a))₂,—N(R^(a))C(NR^(a))N(R^(a))₂, —N(R^(a))C(═O)R^(a),—N(R^(a))S(═O)_(t)R^(a) (where t is 1 or 2), —N(R^(a))S(═O)_(t)N(R^(a))₂(where t is 1 or 2), —S(═O)_(t)R^(a) (where t is 1 or 2),—S(═O)_(t)N(R^(a))₂ (where t is 1 or 2), —PO₃(R^(a))₂, OPO₃WY (where Wand Y are independently hydrogen, methyl, ethyl, alkyl, lithium, sodiumor potassium) or —OPO₃Z (where Z is calcium, magnesium or iron), whereineach R^(a) is independently hydrogen, alkyl, fluoroalkyl, heteroalkyl,cycloalkyl, heterocycloalkyl, heterocycloalkylalkyl, aryl, aralkyl,heteroaryl or heteroarylalkyl.

The term “fluoroalkyl” refers to an alkyl group substituted with one ormore fluorine atoms. In some embodiments, it is a C₁-C₄ alkyl groupsubstituted with one or more fluorine atoms. Typical fluoroalkyl groupsinclude, but are in no way limited to, —CH₂F, —CHF₂, —CF₃, —CH₂CF₃,—CH₂CHF₂, —CH₂CH₂F, —CHFCH₃ and —CF₂CH₃.

The term “alkenyl” refers to a straight or branched hydrocarbon chainradical group comprising carbon and hydrogen atoms, containing at leastone double bond, and having from two to ten carbon atoms (i.e., C₂-C₁₀alkenyl). Whenever it appears herein, a numerical range such as “2 to10” refers to each integer in the given range; e.g., “2 to 10 carbonatoms” means that the alkenyl group may contain 2 carbon atoms, 3 carbonatoms, etc., up to and including 10 carbon atoms. In certainembodiments, an alkenyl comprises two to eight carbon atoms (i.e., C₂-C₈alkenyl). In other embodiments, an alkenyl comprises two to five carbonatoms (i.e., C₂-C₅ alkenyl). The alkenyl is attached to the rest of themolecule by a single bond, for example, ethenyl (i.e., vinyl),prop-1-enyl, but-1-enyl, pent-1-enyl, penta-1,4-dienyl, and the like.Unless stated otherwise specifically in the specification, an alkenylgroup is optionally substituted by one or more of the followingsubstituents: alkyl, heteroalkyl, alkenyl, alkynyl, cycloalkyl,heterocycloalkyl, aryl, arylalkyl, heteroaryl, heteroarylalkyl, hydroxy,halo, cyano, trifluoromethyl, trifluoromethoxy, nitro, oxo, thioxo,trimethylsilanyl, —OR^(a), —SR^(a), —OC(═O)R^(a), —OC(═O)OR^(a),—OC(═O)N(R^(a))₂, —N(R^(a))₂, —C(═O)OR^(a), —C(═O)R^(a),—C(═O)N(R^(a))₂, —N(R^(a))C(═O)OR^(a), —N(R^(a))C(═O)N(R^(a))₂,—N(R^(a))C(NR^(a))N(R^(a))₂, —N(R^(a))C(═O)R^(a),—N(R^(a))S(═O)_(t)R^(a) (where t is 1 or 2), —N(R^(a))S(═O)_(t)N(R^(a))₂(where t is 1 or 2), —S(═O)_(t)R^(a) (where t is 1 or 2),—S(═O)_(t)N(R^(a))₂ (where t is 1 or 2), —PO₃(R^(a))₂, —OPO₃WY (where Wand Y are independently hydrogen, methyl, ethyl, alkyl, lithium, sodiumor potassium) or —OPO₃Z (where Z is calcium, magnesium or iron), whereineach R^(a) is independently hydrogen, alkyl, fluoroalkyl, heteroalkyl,cycloalkyl, heterocycloalkyl, heterocycloalkylalkyl, aryl, aralkyl,heteroaryl or heteroarylalkyl.

The term “alkynyl” refers to a straight or branched hydrocarbon chainradical group comprising carbon and hydrogen atoms, containing at leastone triple bond, and having from two to ten carbon atoms (i.e., C₂-C₁₀alkynyl). In some embodiments, an alkynyl group may contain one or moredouble bonds. Whenever it appears herein, a numerical range such as “2to 10” refers to each integer in the given range; e.g., “2 to 10 carbonatoms” means that the alkynyl group may contain 2 carbon atoms, 3 carbonatoms, etc., up to and including 10 carbon atoms. In certainembodiments, an alkynyl comprises two to eight carbon atoms (i.e., C₂-C₈alkynyl). In other embodiments, an alkynyl has two to five carbon atoms(i.e., C₂-C₅ alkynyl). The alkynyl is attached to the rest of themolecule by a single bond, for example, ethynyl, propynyl, butynyl,pentynyl, hexynyl, and the like. Unless stated otherwise specifically inthe specification, an alkynyl group is optionally substituted by one ormore of the following substituents: alkyl, heteroalkyl, alkenyl,alkynyl, cycloalkyl, heterocycloalkyl, aryl, arylalkyl, heteroaryl,heteroarylalkyl, hydroxy, halo, cyano, trifluoromethyl,trifluoromethoxy, nitro, oxo, thioxo, trimethylsilanyl, —OR^(a),—SR^(a), —OC(═O)R^(a), —OC(═O)OR^(a), —OC(═O)N(R^(a))₂, —N(R^(a))₂,—C(═O)OR^(a), —C(═O)R^(a), —C(═O)N(R^(a))₂, —N(R^(a))C(═O)OR^(a),—N(R^(a))C(═O)N(R^(a))₂, —N(R^(a))C(NR^(a))N(R^(a))₂,—N(R^(a))C(═O)R^(a), —N(R^(a))S(═O)_(t)R^(a) (where t is 1 or 2),—N(R^(a))S(═O)_(t)N(R^(a))₂ (where t is 1 or 2), —S(═O)_(t)R^(a) (wheret is 1 or 2), —S(═O)_(t)N(R^(a))₂ (where t is 1 or 2), —PO₃(R^(a))₂,—OPO₃WY (where W and Y are independently hydrogen, methyl, ethyl, alkyl,lithium, sodium or potassium) or —OPO₃Z (where Z is calcium, magnesiumor iron), wherein each R^(a) is independently hydrogen, alkyl,fluoroalkyl, heteroalkyl, cycloalkyl, heterocycloalkyl,heterocycloalkylalkyl, aryl, aralkyl, heteroaryl or heteroarylalkyl.

The term “aromatic” or “aryl” refers to an aromatic radical with six toten ring atoms (i.e., C₆-C₁₀ aromatic or C₆-C₁₀ aryl) which has at leastone ring having a conjugated pi electron system which is carbocyclic(e.g., phenyl, fluorenyl, and naphthyl). Whenever it appears herein, anumerical range such as “6 to 10” refers to each integer in the givenrange; e.g., “6 to 10 ring atoms” means that the aryl group may consistof 6 ring atoms, 7 ring atoms, etc., up to and including 10 ring atoms.The term includes monocyclic or fused-ring polycyclic (i.e., rings whichshare adjacent pairs of ring atoms) groups. Unless stated otherwisespecifically in the specification, an aryl moiety is optionallysubstituted by one or more substituents which are independently: alkyl,heteroalkyl, alkenyl, alkynyl, cycloalkyl, heterocycloalkyl, aryl,arylalkyl, heteroaryl, heteroarylalkyl, hydroxy, halo, cyano,trifluoromethyl, trifluoromethoxy, nitro, oxo, thioxo, trimethylsilanyl,—OR^(a), —SR^(a), —OC(═O)R^(a), —OC(═O)OR^(a), —OC(═O)N(R^(a))₂,—N(R^(a))₂, —C(═O)OR^(a), —C(═O)R^(a), —C(═O)N(R^(a))₂,—N(R^(a))C(═O)OR^(a), —N(R^(a))C(═O)N(R^(a))₂,—N(R^(a))C(NR^(a))N(R^(a))₂, —N(R^(a))C(═O)R^(a),—N(R^(a))S(═O)_(t)R^(a) (where t is 1 or 2), —N(R^(a))S(═O)_(t)N(R^(a))₂(where t is 1 or 2), —S(═O)_(t)R^(a) (where t is 1 or 2),—S(═O)_(t)N(R^(a))₂ (where t is 1 or 2), —PO₃(R^(a))₂, —OPO₃WY (where Wand Y are independently hydrogen, methyl, ethyl, alkyl, lithium, sodiumor potassium) or —OPO₃Z (where Z is calcium, magnesium or iron), whereineach R^(a) is independently hydrogen, alkyl, fluoroalkyl, heteroalkyl,cycloalkyl, heterocycloalkyl, heterocycloalkylalkyl, aryl, aralkyl,heteroaryl or heteroarylalkyl.

“Aralkyl” or “arylalkyl” refers to an (aryl)alkyl-radical wherein thearylalkyl moiety is attached via the alkyl portion of the moiety. Aryland alkyl are as disclosed herein and are optionally substituted by oneor more of the substituents described as suitable substituents for aryland alkyl, respectively.

The term “heteroaryl” or, alternatively, “heteroaromatic” refers to a 5-to 18-membered aromatic radical (i.e., C₅-C₁₈ heteroaryl) that includesone or more ring heteroatoms selected from nitrogen, oxygen and sulfur,and which may be a monocyclic, bicyclic, tricyclic or tetracyclic ringsystem. Whenever it appears herein, a numerical range such as “5 to 18”refers to each integer in the given range; e.g., “5 to 18 ring atoms”means that the heteroaryl group may consist of 5 ring atoms, 6 ringatoms, etc., up to and including 18 ring atoms. An N-containing“heteroaromatic” or “heteroaryl” moiety refers to an aromatic group inwhich at least one of the skeletal atoms of the ring is a nitrogen atom.The polycyclic heteroaryl group may be fused or non-fused. Theheteroatom(s) in the heteroaryl radical, e.g., nitrogen or sulfur, isoptionally oxidized. One or more nitrogen atoms, if present, areoptionally quaternized. The heteroaryl is attached to the rest of themolecule through any atom of the ring(s). Examples of heteroarylsinclude, but are not limited to, azepinyl, acridinyl, benzimidazolyl,benzindolyl, 1,3-benzodioxolyl, benzooxazolyl, benzo[d]thiazolyl,benzothiadiazolyl, benzo[b][1,4]dioxepinyl, benzo[b][1,4]oxazinyl,1,4-benzodioxanyl, benzonaphthofuranyl, benzoxazolyl, benzodioxolyl,benzodioxinyl, benzopyranyl, benzopyranonyl, benzofuranyl,benzofuranonyl, benzofurazanyl, benzothiazolyl, benzothienyl,benzothieno[3,2-d]pyrimidinyl, benzotriazolyl,benzo[4,6]imidazo[1,2-a]pyridinyl, carbazolyl, cinnolinyl,cyclopenta[d]pyrimidinyl,6,7-dihydro-5H-cyclopenta[4,5]thieno[2,3-d]pyrimidinyl,5,6-dihydrobenzo[h]quinazolinyl, 5,6-dihydrobenzo[h]cinnolinyl,6,7-dihydro-5H-benzo[6,7]cyclohepta[1,2-c]pyridazinyl, dibenzofuranyl,dibenzothiophenyl, furanyl, furazanyl, furanonyl, furo[3,2-c]pyridinyl,5,6,7,8,9,10-hexahydrocycloocta[d]pyrimidinyl,5,6,7,8,9,10-hexahydrocycloocta[d]pyridazinyl,5,6,7,8,9,10-hexahydrocycloocta[d]pyridinyl, isothiazolyl, imidazolyl,indazolyl, indolyl, indazolyl, isoindolyl, indolinyl, isoindolinyl,isoquinolyl, indolizinyl, isoxazolyl,5,8-methano-5,6,7,8-tetrahydroquinazolinyl, naphthyridinyl,1,6-naphthyridinonyl, oxadiazolyl, 2-oxoazepinyl, oxazolyl, oxiranyl,5,6,6a,7,8,9,10,10a-octahydrobenzo[h]quinazolinyl, 1-phenyl-1H-pyrrolyl,phenazinyl, phenothiazinyl, phenoxazinyl, phthalazinyl, pteridinyl,purinyl, pyranyl, pyrrolyl, pyrazolyl, pyrazolo[3,4-d]pyrimidinyl,pyridinyl, pyrido[3,2-d]pyrimidinyl, pyrido[3,4-d]pyrimidinyl,pyrazinyl, pyrimidinyl, pyridazinyl, pyrrolyl, quinazolinyl,quinoxalinyl, quinolinyl, isoquinolinyl, tetrahydroquinolinyl,5,6,7,8-tetrahydroquinazolinyl,5,6,7,8-tetrahydrobenzo[4,5]thieno[2,3-d]pyrimidinyl,6,7,8,9-tetrahydro-5H-cyclohepta[4,5]thieno[2,3-d]pyrimidinyl,5,6,7,8-tetrahydropyrido[4,5-c]pyridazinyl, thiazolyl, thiadiazolyl,thiapyranyl, triazolyl, tetrazolyl, triazinyl, thieno[2,3-d]pyrimidinyl,thieno[3,2-d]pyrimidinyl, thieno[2,3-c]pridinyl, and thiophenyl (i.e.thienyl). Unless stated otherwise specifically in the specification, aheteroaryl moiety is optionally substituted by one or more substituentswhich are independently alkyl, heteroalkyl, alkenyl, alkynyl,cycloalkyl, heterocycloalkyl, aryl, arylalkyl, heteroaryl,heteroarylalkyl, hydroxy, halo, cyano, trifluoromethyl,trifluoromethoxy, nitro, oxo, thioxo, trimethylsilanyl, —OR^(a),—SR^(a), —OC(═O)R^(a), —OC(═O)OR^(a), —OC(═O)N(R^(a))₂, —N(R^(a))₂,—C(═O)OR^(a), —C(═O)R^(a), —C(═O)N(R^(a))₂, —N(R^(a))C(═O)OR^(a),—N(R^(a))C(═O)N(R^(a))₂, —N(R^(a))C(NR^(a))N(R^(a))₂,—N(R^(a))C(═O)R^(a), —N(R^(a))S(═O)_(t)R^(a) (where t is 1 or 2),—N(R^(a))S(═O)_(t)N(R^(a))₂ (where t is 1 or 2), —S(═O)_(t)R^(a) (wheret is 1 or 2), —S(═O)_(t)N(R^(a))₂ (where t is 1 or 2), —PO₃(R^(a))₂,—OPO₃WY (where W and Y are independently hydrogen, methyl, ethyl, alkyl,lithium, sodium or potassium) or —OPO₃Z (where Z is calcium, magnesiumor iron), wherein each R^(a) is independently hydrogen, alkyl,fluoroalkyl, heteroalkyl, cycloalkyl, heterocycloalkyl,heterocycloalkylalkyl, aryl, aralkyl, heteroaryl or heteroarylalkyl.Examples of monocylic heteroaryls include, but are not limited to,imidazolyl, pyridinyl, pyrrolyl, pyrazinyl, pyrimidinyl, thiazolyl,furanyl and thienyl.

Substituted heteroaryl also includes ring systems substituted with oneor more oxide substituents, such as pyridinyl N-oxides.

“Heteroarylalkyl” refers to a moiety having a heteroaryl moiety, asdescribed herein, connected to an alkyl moiety, as described herein,wherein the connection to the remainder of the molecule is through thealkyl group. Heteroaryl and alkyl are as disclosed herein and areoptionally substituted by one or more of the substituents described assuitable substituents for heteroaryl and alkyl, respectively.

The term “acyl” refers to a —C(═O)R radical, wherein R is alkyl,cycloalkyl, aryl, heteroaryl, heteroalkyl, or heterocycloalkyl, whichare as described herein. The R group is attached to the parent structurethrough the carbonyl functionality. In some embodiments, it is a C₁-C₁₀acyl radical which refers to the total number of chain or ring atoms ofthe alkyl, cycloalkyl, aryl, heteroalkyl, heteroaryl or heterocycloalkylportion of the acyl group plus the carbonyl carbon of acyl, i.e. ring orchain atoms plus carbonyl. If the R radical is heteroaryl orheterocycloalkyl, the hetero ring or chain atoms contribute to the totalnumber of chain or ring atoms. Unless stated otherwise specifically inthe specification, the “R” of an acyl group is optionally substituted byone or more substituents which independently are: alkyl, heteroalkyl,alkenyl, alkynyl, cycloalkyl, heterocycloalkyl, aryl, arylalkyl,heteroaryl, heteroarylalkyl, hydroxy, halo, cyano, trifluoromethyl,trifluoromethoxy, nitro, oxo, thioxo, trimethylsilanyl, —OR^(a),—SR^(a), —OC(═O)R^(a), —OC(═O)OR^(a), —OC(═O)N(R^(a))₂, —N(R^(a))₂,—C(═O)OR^(a), —C(═O)R^(a), —C(═O)N(R^(a))₂, —N(R^(a))C(═O)OR^(a),—N(R^(a))C(═O)N(R^(a))₂, —N(R^(a))C(NR^(a))N(R^(a))₂,—N(R^(a))C(═O)R^(a), —N(R^(a))S(═O)_(t)R^(a) (where t is 1 or 2),—N(R^(a))S(═O)_(t)N(R^(a))₂ (where t is 1 or 2), —S(═O)_(t)R^(a) (wheret is 1 or 2), —S(═O)_(t)N(R^(a))₂ (where t is 1 or 2), —PO₃(R^(a))₂,—OPO₃WY (where W and Y are independently hydrogen, methyl, ethyl, alkyl,lithium, sodium or potassium) or —OPO₃Z (where Z is calcium, magnesiumor iron), wherein each R^(a) is independently hydrogen, alkyl,fluoroalkyl, heteroalkyl, cycloalkyl, heterocycloalkyl,heterocycloalkylalkyl, aryl, aralkyl, heteroaryl or heteroarylalkyl.

The term “halo”, “halide”, or alternatively, “halogen” means fluoro,chloro, bromo or iodo. The terms “haloalkyl,” “haloalkenyl,”“haloalkynyl” and “haloalkoxy” include alkyl, alkenyl, alkynyl andalkoxy structures that are substituted with one or more halo groups orwith combinations thereof. For example, the terms “fluoroalkyl” and“fluoroalkoxy” refer to haloalkyl and haloalkoxy groups, respectively,in which the halo is fluoro. Examples of fluoroalkyl include, but arenot limited to, —CH₂F, —CHF₂, —CF₃, —CF₂CH₃, —CH₂CF₃, and —CF₂CF₃. Thealkyl part of the haloalkyl radical may be optionally substituted asdefined above for an alkyl group.

The term “cyano” refers to a —CN radical.

The term “alkoxy” refers to an —O-alkyl radical, including from whereinalkyl is as described herein and contains 1 to 10 carbon atoms (i.e.,C₁-C₁₀ alkoxy) of a straight, branched, or cyclic configuration andcombinations thereof attached to the parent structure through an oxygen.Examples include methoxy, ethoxy, propoxy, isopropoxy, cyclopropyloxy,cyclohexyloxy and the like. Whenever it appears herein, a numericalrange such as “1 to 10” refers to each integer in the given range; e.g.,“1 to 10 carbon atoms” means that the alkyl group may consist of 1carbon atom, 2 carbon atoms, 3 carbon atoms, etc., up to and including10 carbon atoms. In some embodiments, it is a C₁-C₄ alkoxy group. Unlessstated otherwise specifically in the specification, an alkoxy moiety maybe substituted by one or more of the substituents described as suitablesubstituents for an alkyl radical.

The term “sp³ hybridized carbon” refers to a carbon atom that is bondedto four other atoms. sp³ hybridization results from the combination ofthe s orbital and all three p orbitals in the second energy level ofcarbon. It results in four equivalent orbitals and the geometricarrangement of those four orbitals is tetrahedral.

The term “sulfonyl” refers to a —S(═O)₂R^(a) radical, wherein R^(a) isselected from the group consisting of alkyl, amino, cycloalkyl, aryl,heteroalkyl, heteroaryl (bonded through a ring carbon) andheterocycloalkyl (bonded through a ring carbon). Unless stated otherwisespecifically in the specification, the R^(a) group may be substituted byone or more of the substituents described as suitable substituents foran alkyl, an aryl or a heteroaryl radical.

The term “sulfoximinyl” refers to a —S(═O)(═NR^(a))R^(b) radical,wherein R^(a) is selected from the group consisting of hydrogen, alkyl,cycloalkyl, aryl, cyano, carbamoyl, acyl, heteroaryl (bonded through aring carbon) and heterocycloalkyl (bonded through a ring carbon) andR^(b) is independently selected from the group consisting of alkyl,cycloalkyl, aryl, heteroalkyl, heteroaryl (bonded through a ring carbon)and heterocycloalkyl (bonded through a ring carbon). Unless statedotherwise specifically in the specification, the R^(a) and R^(b) groupsmay be substituted by one or more of the substituents described assuitable substituents for an alkyl, an aryl or a heteroaryl radical.

“Sulfonamide,” “sulfonamidyl” or “sulfonamido” refers to a—S(═O)₂N(R^(a))₂ radical, wherein each R^(a) is selected independentlyfrom the group consisting of hydrogen, alkyl, heteroalkyl, cycloalkyl,aryl, heteroaryl and heterocycloalkyl. The R^(a) groups in —N(R^(a))₂ ofthe —S(═O)₂—N(R^(a))₂ radical may be taken together with the nitrogen towhich it is attached to form a 4-, 5-, 6-, or 7-membered ring. In someembodiments, it is a C₁-C₁₀ sulfonamido, wherein each R^(a) insulfonamido contains 1 carbon, 2 carbons, 3 carbons or 4 carbons total.A sulfonamido group is optionally substituted by one or more of thesubstituents described for alkyl, cycloalkyl, aryl and heteroaryl,respectively.

The term “fluoroalkylsulfonyl” refers to a —S(═O)₂R^(a) radical, whereinR^(a) is fluoroalkyl. In some embodiments, R^(a) is C₁-C₄ alkyl,substituted with one or more fluorines.

The term “cycloalkyl” refers to a monocyclic or polycyclic non-aromaticradical that contains carbon and hydrogen, and may be saturated, orpartially unsaturated. Cycloalkyl groups include groups having from 3 to10 ring atoms (e.g., C₃-C₁₀ cycloalkyl). Whenever it appears herein, anumerical range such as “3 to 10” refers to each integer in the givenrange; e.g., “3 to 10 carbon atoms” means that the cycloalkyl group mayconsist of 3 carbon ring atoms, 4 carbon ring atoms, 5 carbon ringatoms, etc., up to and including 10 carbon ring atoms. In someembodiments, it is a C₃-C₈ cycloalkyl radical. In some embodiments, itis a C₃-C₅ cycloalkyl radical. Illustrative examples of cycloalkylgroups include, but are not limited to the following moieties:cyclopropyl, cyclobutyl, cyclopentyl, cyclopentenyl, cyclohexyl,cyclohexenyl, cycloseptyl, cyclooctyl, cyclononyl, cyclodecyl,norbornyl, and the like. Unless stated otherwise specifically in thespecification, a cycloalkyl group is optionally substituted by one ormore substituents which independently are: alkyl, heteroalkyl, alkenyl,alkynyl, cycloalkyl, heterocycloalkyl, aryl, arylalkyl, heteroaryl,heteroarylalkyl, hydroxy, halo, cyano, trifluoromethyl,trifluoromethoxy, nitro, oxo, thioxo, trimethylsilanyl, —OR^(a),—SR^(a), —OC(═O)R^(a), —OC(═O)OR^(a), —OC(═O)N(R^(a))₂, —N(R^(a))₂,—C(═O)OR^(a), —C(═O)R^(a), —C(═O)N(R^(a))₂, —N(R^(a))C(═O)OR^(a),—N(R^(a))C(═O)N(R^(a))₂, —N(R^(a))C(NR^(a))N(R^(a))₂,—N(R^(a))C(═O)R^(a), —N(R^(a))S(═O)_(t)R^(a) (where t is 1 or 2),—N(R^(a))S(═O)_(t)N(R^(a))₂ (where t is 1 or 2), —S(═O)_(t)R^(a) (wheret is 1 or 2), —S(═O)_(t)N(R^(a))₂ (where t is 1 or 2), —PO₃(R^(a))₂,—OPO₃WY (where W and Y are independently hydrogen, methyl, ethyl, alkyl,lithium, sodium or potassium) or —OPO₃Z (where Z is calcium, magnesiumor iron), wherein each R^(a) is independently hydrogen, alkyl,fluoroalkyl, heteroalkyl, cycloalkyl, heterocycloalkyl,heterocycloalkylalkyl, aryl, aralkyl, heteroaryl or heteroarylalkyl.

The term “heterocyclyl” or “heterocycloalkyl” refers to a stable 3- to18-membered nonaromatic ring (e.g., C₃-C₁₈ heterocycloalkyl) radicalthat comprises two to twelve ring carbon atoms and from one to sixheteroatoms selected from nitrogen, oxygen and sulfur. Whenever itappears herein, a numerical range such as “3 to 18” refers to eachinteger in the given range; e.g., “3 to 18 ring atoms” means that theheterocycloalkyl group may consist of 3 ring atoms, 4 ring atoms, etc.,up to and including 18 ring atoms. In some embodiments, it is a C₅-C₁₀heterocycloalkyl. In some embodiments, it is a C₄-C₁₀ heterocycloalkyl.In some embodiments, it is a C₃-C₁₀ heterocycloalkyl. Unless statedotherwise specifically in the specification, the heterocycloalkylradical may be a monocyclic, bicyclic, tricyclic or tetracyclic ringsystem, which may include fused or bridged ring systems. The heteroatomsin the heterocycloalkyl radical may be optionally oxidized. One or morenitrogen atoms, if present, may optionally be quaternized. Theheterocycloalkyl radical may be partially or fully saturated. Theheterocycloalkyl may be attached to the rest of the molecule through anyatom of the ring(s). Examples of such heterocycloalkyl radicals include,but are not limited to, 6,7-dihydro-5H-cyclopenta[b]pyridine,dioxolanyl, thienyl[1,3]dithianyl, decahydroisoquinolyl, imidazolinyl,imidazolidinyl, isothiazolidinyl, isoxazolidinyl, morpholinyl,octahydroindolyl, octahydroisoindolyl, 2-oxopiperazinyl,2-oxopiperidinyl, 2-oxopyrrolidinyl, oxazolidinyl, piperidinyl,piperazinyl, 4-piperidonyl, pyrrolidinyl, pyrazolidinyl, quinuclidinyl,thiazolidinyl, tetrahydrofuryl, trithianyl, tetrahydropyranyl,thiomorpholinyl, thiamorpholinyl, 1-oxo-thiomorpholinyl, and1,1-dioxo-thiomorpholinyl. Unless stated otherwise specifically in thespecification, a heterocycloalkyl moiety is optionally substituted byone or more substituents which independently are: alkyl, heteroalkyl,alkenyl, alkynyl, cycloalkyl, heterocycloalkyl, aryl, arylalkyl,heteroaryl, heteroarylalkyl, hydroxy, halo, cyano, trifluoromethyl,trifluoromethoxy, nitro, oxo, thioxo, trimethylsilanyl, —OR^(a),—SR^(a), —OC(═O)R^(a), —OC(═O)OR^(a), —OC(═O)N(R^(a))₂, —N(R^(a))₂,—C(═O)OR^(a), —C(═O)R^(a), —C(═O)N(R^(a))₂, —N(R^(a))C(═O)OR^(a),—N(R^(a))C(═O)N(R^(a))₂, —N(R^(a))C(NR^(a))N(R^(a))₂,—N(R^(a))C(═O)R^(a), —N(R^(a))S(═O)_(t)R^(a) (where t is 1 or 2),—N(R^(a))S(═O)_(t)N(R^(a))₂ (where t is 1 or 2), —S(═O)_(t)R^(a) (wheret is 1 or 2), —S(═O)_(t)N(R^(a))₂ (where t is 1 or 2), —PO₃(R^(a))₂,—OPO₃WY (where W and Y are independently hydrogen, methyl, ethyl, alkyl,lithium, sodium or potassium) or —OPO₃Z (where Z is calcium, magnesiumor iron), wherein each R^(a) is independently hydrogen, alkyl,fluoroalkyl, heteroalkyl, cycloalkyl, heterocycloalkyl,heterocycloalkylalkyl, aryl, aralkyl, heteroaryl or heteroarylalkyl.

“Heterocycloalkyl” also includes bicyclic ring systems wherein onenon-aromatic ring, usually with 3 to 7 ring atoms, contains at least 2carbon atoms in addition to 1-3 heteroatoms independently selected fromoxygen, sulfur and nitrogen, as well as combinations comprising at leastone of the foregoing heteroatoms; the other ring, usually with 3 to 7ring atoms, optionally contains 1-3 heteroatoms independently selectedform oxygen, sulfur and nitrogen and is not aromatic.

The terms “heteroalkyl”, “heteroalkenyl” and “heteroalkynyl” includeoptionally substituted alkyl, alkenyl and alkynyl radicals, whichrespectively have one or more skeletal chain atoms selected from an atomother than carbon, e.g., oxygen, nitrogen, sulfur, phosphorus orcombinations thereof. A numerical range, which refers to the chainlength in total, may be given. For example, C₃-C₄ heteroalkyl has achain length of 3-4 atoms. For example, a —CH₂OCH₂CH₃ radical isreferred to as a “C₄ heteroalkyl”, which includes the heteroatom in theatom chain length description. Connection to the rest of the moleculemay be through either a heteroatom or a carbon in the heteroalkyl chain.A heteroalkyl may be a substituted alkyl. The same definition applies toheteroalkenyl or heteroalkynyl. Unless otherwise stated in thespecification, a heteroalkyl group may be substituted with one or moresubstituents which independently are: alkyl, heteroalkyl, alkenyl,alkynyl, cycloalkyl, heterocycloalkyl, aryl, arylalkyl, heteroaryl,heteroarylalkyl, hydroxy, halo, cyano, trifluoromethyl,trifluoromethoxy, nitro, oxo, thioxo, trimethylsilanyl, —OR^(a),—SR^(a), —OC(═O)R^(a), —OC(═O)OR^(a), —OC(═O)N(R^(a))₂, —N(R^(a))₂,—C(═O)OR^(a), —C(═O)R^(a), —C(═O)N(R^(a))₂, —N(R^(a))C(═O)OR^(a),—N(R^(a))C(═O)N(R^(a))₂, —N(R^(a))C(NR^(a))N(R^(a))₂,—N(R^(a))C(═O)R^(a), —N(R^(a))S(═O)_(t)R^(a) (where t is 1 or 2),—N(R^(a))S(═O)_(t)N(R^(a))₂ (where t is 1 or 2), —S(═O)_(t)R^(a) (wheret is 1 or 2), —S(═O)_(t)N(R^(a))₂ (where t is 1 or 2), —PO₃(R^(a))₂,—OPO₃WY (where W and Y are independently hydrogen, methyl, ethyl, alkyl,lithium, sodium or potassium) or —OPO₃Z (where Z is calcium, magnesiumor iron), wherein each R^(a) is independently hydrogen, alkyl,fluoroalkyl, heteroalkyl, cycloalkyl, heterocycloalkyl,heterocycloalkylalkyl, aryl, aralkyl, heteroaryl or heteroarylalkyl.

The term “amino” or “amine” refers to a —N(R^(a))₂ radical group, whereeach R^(a) is independently hydrogen, alkyl, heteroalkyl, fluoroalkyl,cycloalkyl, aryl, aralkyl, heterocycloalkyl, heterocycloalkylalkyl,heteroaryl or heteroarylalkyl, unless stated otherwise specifically inthe specification. When a —N(R^(a))₂ group has two R^(a) other thanhydrogen, they can be combined with the nitrogen atom to form a 3-, 4-,5-, 6-, 7- or 8-membered ring. For example, —N(R^(a))₂ is meant toinclude, but not be limited to, 1-pyrrolidinyl and 4-morpholinyl. Unlessstated otherwise specifically in the specification, an amino group isoptionally substituted by one or more substituents which independentlyare: alkyl, heteroalkyl, alkenyl, alkynyl, cycloalkyl, heterocycloalkyl,aryl, arylalkyl, heteroaryl, heteroarylalkyl, hydroxy, halo, cyano,trifluoromethyl, trifluoromethoxy, nitro, oxo, thioxo, trimethylsilanyl,—OR^(a), —SR^(a), —OC(═O)R^(a), —OC(═O)OR^(a), —OC(═O)N(R^(a))₂,—N(R^(a))₂, —C(═O)OR^(a), —C(═O)R^(a), —C(═O)N(R^(a))₂,—N(R^(a))C(═O)OR^(a), —N(R^(a))C(═O)N(R^(a))₂,—N(R^(a))C(NR^(a))N(R^(a))₂, —N(R^(a))C(═O)R^(a),—N(R^(a))S(═O)_(t)R^(a) (where t is 1 or 2), —N(R^(a))S(═O)_(t)N(R^(a))₂(where t is 1 or 2), —S(═O)_(t)R^(a) (where t is 1 or 2),—S(═O)_(t)N(R^(a))₂ (where t is 1 or 2), —PO₃(R^(a))₂, OPO₃WY (where Wand Y are independently hydrogen, methyl, ethyl, alkyl, lithium, sodiumor potassium) or —OPO₃Z (where Z is calcium, magnesium or iron), whereineach R^(a) is independently hydrogen, alkyl, fluoroalkyl, heteroalkyl,cycloalkyl, heterocycloalkyl, heterocycloalkylalkyl, aryl, aralkyl,heteroaryl or heteroarylalkyl.

The term “substituted amino” also refers to N-oxides of the groupN(R^(a))₂ as described above. N-oxides can be prepared by treatment ofthe corresponding amino group with, for example, hydrogen peroxide orm-chloroperoxybenzoic acid. The person skilled in the art is familiarwith reaction conditions for carrying out the N-oxidation.

The term “acyloxy” refers to a RC(═O)O— radical wherein R is alkyl,cycloalkyl, aryl, heteroalkyl, heteroaryl or heterocycloalkyl, which areas described herein. In some embodiments, it is a C₁-C₄ acyloxy radical,which refers to the total number of chain or ring atoms of the alkyl,cycloalkyl, aryl, heteroalkyl, heteroaryl or heterocycloalkyl portion ofthe acyloxy group plus the carbonyl carbon of acyl, i.e., the other ringor chain atoms plus carbonyl. If the R radical is heteroaryl orheterocycloalkyl, the hetero ring or chain atoms contribute to the totalnumber of chain or ring atoms. Unless stated otherwise specifically inthe specification, the “R” of an acyloxy group is optionally substitutedby one or more of the following substituents: alkyl, heteroalkyl,alkenyl, alkynyl, cycloalkyl, heterocycloalkyl, aryl, arylalkyl,heteroaryl, heteroarylalkyl, hydroxy, halo, cyano, trifluoromethyl,trifluoromethoxy, nitro, oxo, thioxo, trimethylsilanyl, —OR^(a),—SR^(a), —OC(═O)R^(a), —OC(═O)OR^(a), —OC(═O)N(R^(a))₂, —N(R^(a))₂,—C(═O)OR^(a), —C(═O)R^(a), —C(═O)N(R^(a))₂, —N(R^(a))C(═O)OR^(a),—N(R^(a))C(═O)N(R^(a))₂, —N(R^(a))C(NR^(a))N(R^(a))₂,—N(R^(a))C(═O)R^(a), —N(R^(a))S(═O)_(t)R^(a) (where t is 1 or 2),—N(R^(a))S(═O)_(t)N(R^(a))₂ (where t is 1 or 2), —S(═O)_(t)R^(a) (wheret is 1 or 2), —S(═O)_(t)N(R^(a))₂ (where t is 1 or 2), —PO₃(R^(a))₂,—OPO₃WY (where W and Y are independently hydrogen, methyl, ethyl, alkyl,lithium, sodium or potassium) or —OPO₃Z (where Z is calcium, magnesiumor iron), wherein each R^(a) is independently hydrogen, alkyl,fluoroalkyl, heteroalkyl, cycloalkyl, heterocycloalkyl,heterocycloalkylalkyl, aryl, aralkyl, heteroaryl or heteroarylalkyl.

The term “amide” or “amido” refers to a chemical moiety with formula—C(═O)N(R^(a))₂ or —NR^(a)C(═O)R^(a), wherein each of R^(a) isindependently selected from the group consisting of hydrogen, alkyl,cycloalkyl, aryl, heteroaryl (bonded through a ring carbon) andheterocycloalkyl. Two R^(a)s may optionally be taken together with thenitrogen to which it is attached to form a 4-10 membered ring. In someembodiments, it is a C₁-C₄ amido or amide radical, which includes theamide carbonyl in the total number of carbons in the radical. Unlessstated otherwise specifically in the specification, an amido group isoptionally substituted independently by one or more of the substituentsas described herein for alkyl, cycloalkyl, aryl, heteroaryl, orheterocycloalkyl. An amide may be an amino acid or a peptide moleculeattached to a compound having an amine or a carboxylic acid moiety,thereby forming a prodrug. Any amine, hydroxy or carboxyl side chain onthe compounds described herein can be amidified. The procedures andspecific groups to make such amides are known to those of skilled in theart and can readily be found in reference sources such as Wuts, Greene'sProtective Groups in Organic Synthesis, 5^(th) Ed., Wiley, New York,N.Y., 2014, which is incorporated herein by reference in its entirety.

“Carboxaldehyde” refers to a —C(═O)H radical.

“Carboxyl” refers to a —C(═O)OH radical.

“Ester” refers to a chemical radical of formula —C(═O)OR, where R isselected from the group consisting of alkyl, cycloalkyl, aryl,heteroaryl (bonded through a ring carbon) and heterocycloalkyl (bondedthrough a ring carbon). Any amine, hydroxy, or carboxyl side chain onthe compounds described herein can be esterified. The procedures andspecific groups to make such esters are known to those skilled in theart and can readily be found in reference sources such as Wuts, Greene'sProtective Groups in Organic Synthesis, 5^(th) Ed., Wiley, New York,N.Y., 2014, which is incorporated herein by reference in its entirety.Unless stated otherwise specifically in the specification, an estergroup is optionally substituted by one or more substituents whichindependently are: alkyl, heteroalkyl, alkenyl, alkynyl, cycloalkyl,heterocycloalkyl, aryl, arylalkyl, heteroaryl, heteroarylalkyl, hydroxy,halo, cyano, trifluoromethyl, trifluoromethoxy, nitro, oxo, thioxo,trimethylsilanyl, —OR^(a), —SR^(a), —OC(═O)R^(a), —OC(═O)OR^(a),—OC(═O)N(R^(a))₂, —N(R^(a))₂, —C(═O)OR^(a), —C(═O)R^(a),—C(═O)N(R^(a))₂, —N(R^(a))C(═O)OR^(a), —N(R^(a))C(═O)N(R^(a))₂,—N(R^(a))C(NR^(a))N(R^(a))₂, —N(R^(a))C(═O)R^(a),—N(R^(a))S(═O)_(t)R^(a) (where t is 1 or 2), —N(R^(a))S(═O)_(t)N(R^(a))₂(where t is 1 or 2), —S(═O)_(t)R^(a) (where t is 1 or 2),—S(═O)_(t)N(R^(a))₂ (where t is 1 or 2), —PO₃(R^(a))₂, —OPO₃WY (where Wand Y are independently hydrogen, methyl, ethyl, alkyl, lithium, sodiumor potassium) or —OPO₃Z (where Z is calcium, magnesium or iron), whereineach R^(a) is independently hydrogen, alkyl, fluoroalkyl, heteroalkyl,cycloalkyl, heterocycloalkyl, heterocycloalkylalkyl, aryl, aralkyl,heteroaryl or heteroarylalkyl.

“Imino” refers to a ═N—R^(a) radical, wherein R^(a) is hydrogen, alkyl,heteroalkyl, cycloalkyl, cyano, aryl, heterocycloalkyl or heteroaryl.

“Isocyanato” refers to a —NCO radical.

“Isothiocyanato” refers to a —NCS radical.

“Mercaptyl” refers to an —S(alkyl) or —SH radical.

“Methylene” refers to a ═CH₂ radical.

“Hydroxy” refers to a —OH radical.

“Oxa” refers to a —O— radical.

“Oxo” refers to a═O radical.

“Nitro” refers to a —NO₂ radical.

“Oxime” refers to a ═N(—OR) radical, where R is hydrogen or alkyl.

“Sulfinyl” refers to a —S(═O)R radical, where R is selected from thegroup consisting of alkyl, cycloalkyl, aryl, heteroalkyl, heteroaryl(bonded through a ring carbon) and heterocycloalkyl (bonded through aring carbon). In some embodiments, R is fluoroalkyl.

“Sulfoxyl” refers to a —S(═O)₂OH radical.

“Sulfonate” refers to a —S(═O)₂OR radical, where R is selected from thegroup consisting of alkyl, cycloalkyl, aryl, heteroalkyl, heteroaryl(bonded through a ring carbon) and heteroalkyl (bonded through a ringcarbon). The R group is optionally substituted by one or more of thesubstituents described for alkyl, heteroalkyl, cycloalkyl,heterocycloalkyl, aryl, and heteroaryl, respectively.

“Thiocyanato” refers to a —CNS radical.

“Thioxo” refers to a ═S radical.

“Moiety” refers to a specific segment or functional group of a molecule.Chemical moieties are often recognized chemical entities embedded in orappended to a molecule.

“Substituted” means that the referenced group may be substituted withone or more additional group(s) individually and independently selectedfrom acyl, alkyl, alkylaryl, heteroalkyl, cycloalkyl, aralkl,heterocycloalkyl, aryl, carbohydrate, carbonate, heteroaryl, hydroxy,alkoxy, aryloxy, mercapto, alkylthio, arylthio, cyano, halo, carbonyl,ester, thiocarbonyl, isocyanato, thiocyanato, isothiocyanato, nitro,oxo, perhaloalkyl, perfluoroalkyl, phosphate, silyl, sulfinyl, sulfonyl,sulfonamide, sulfoxyl, sulfonate, urea, and amino, including mono- anddi-substituted amino groups and the protected derivatives thereof. Thesubstituents themselves may be substituted, for example, a cycloalkylsubstituent may have a halide substituted at one or more ring carbons,and the like. The protecting groups that may form the protectivederivatives of the above substituents are known to those of skill in theart and may be found in references such as Wuts, Greene's ProtectiveGroups in Organic Synthesis, 5^(th) Ed., Wiley, New York, N.Y., 2014.

The term “optional” or “optionally” means that the subsequentlydescribed event or circumstance may or may not occur, and includesinstances where the event or circumstance occurs and instances in whichit does not. For example, “alkyl optionally substituted with”encompasses both “alkyl” and “alkyl” substituted with groups as definedherein. It will be understood by those skilled in the art, with respectto any group containing one or more substituents, that such groups arenot intended to introduce any substitution or substitution patternswhich would be deemed unacceptable by one of ordinary skill in the art.

Compounds of the present invention also include crystalline andamorphous forms of those compounds, pharmaceutically acceptable salts,and active metabolites of these compounds having the same type ofactivity, including, for example, polymorphs, pseudopolymorphs,solvates, hydrates, unsolvated polymorphs (including anhydrates),conformational polymorphs, and amorphous forms of the compounds, as wellas mixtures thereof. “Crystalline form,” “polymorph,” and “novel form”may be used interchangeably herein, and are meant to include allcrystalline and amorphous forms of the compound, including, for example,polymorphs, pseudopolymorphs, solvates, hydrates, unsolvated polymorphs(including anhydrates), conformational polymorphs, and amorphous forms,as well as mixtures thereof, unless a particular crystalline oramorphous form is referred to.

The compounds described herein may exhibit their natural isotopicabundance, or one or more of the atoms may be artificially enriched in aparticular isotope having the same atomic number, but an atomic mass ormass number different from the atomic mass or mass number predominantlyfound in nature. All isotopic variations of the compounds of the presentinvention, whether radioactive or not, are encompassed within the scopeof the present invention. For example, hydrogen has three naturallyoccurring isotopes, denoted ¹H (protium), ²H (deuterium), and ³H(tritium). Protium is the most abundant isotope in nature. Enriching fordeuterium may afford certain therapeutic advantages, such as increasedin vivo half-life and/or exposure, or may provide a compound useful forinvestigating in vivo routes of drug elimination and metabolism.Isotopically-enriched compounds may be prepared by conventionaltechniques well known to those skilled in the art or by processesanalogous to those described in the Schemes and Examples herein usingappropriate isotopically-enriched reagents and/or intermediates. SeePleiss and Voger, Synthesis and Applications of Isotopically LabeledCompounds, Vol. 7, Wiley, ISBN-10: 0471495018, published on Mar. 14,2001.

“Isomers” are different compounds that have the same molecular formula.“Stereoisomers” are isomers that differ only in the way the atoms arearranged in space. “Enantiomers” are a pair of stereoisomers that arenon superimposable mirror images of each other. A 1:1 mixture of a pairof enantiomers is a “racemic” mixture. The term “(±)” is used todesignate a racemic mixture where appropriate. “Diastereoisomers” or“diastereomers” are stereoisomers that have at least two asymmetricatoms but are not mirror images of each other. The absolutestereochemistry is specified according to the Cahn-Ingold-Prelog R-Ssystem. When a compound is a pure enantiomer, the stereochemistry ateach chiral carbon can be specified by either R or S. Resolved compoundswhose absolute configuration is unknown can be designated (+) or (−)depending on the direction (dextro- or levorotatory) in which theyrotate plane polarized light at the wavelength of the sodium D line.Certain compounds described herein contain one or more asymmetriccenters and can thus give rise to enantiomers, diastereomers, and otherstereoisomeric forms that can be defined, in terms of absolutestereochemistry, as (R)- or (S)-. The present chemical entities,pharmaceutical compositions and methods are meant to include all suchpossible isomers, including racemic mixtures, optically pure forms,mixtures of diastereomers and intermediate mixtures. Optically active(R)- and (S)-isomers can be prepared using chiral synthons or chiralreagents, or resolved using conventional techniques. The opticalactivity of a compound can be analyzed via any suitable method,including but not limited to chiral chromatography and polarimetry, andthe degree of predominance of one stereoisomer over the other isomer canbe determined.

Chemical entities having carbon-carbon double bonds or carbon-nitrogendouble bonds may exist in Z- or E-form (or cis- or trans-form).Furthermore, some chemical entities may exist in various tautomericforms. Unless otherwise specified, chemical entities described hereinare intended to include all Z-, E- and tautomeric forms as well.

The term “enantiomeric excess,” as used herein, is the percent excess ofone enantiomer compared to that of the other enantiomer in a mixture,and can be calculated using the following equation: enantiomericexcess=((R−S)/(R+S))×100=% (R*)−% (S*), wherein R and S are the numberof moles of each enantiomer in the mixture, and R* and S* are therespective mole fractions of the enantiomers in the mixture. Forexample, for a mixture with 87% R enantiomer and 13% S enantiomer, theenantiomeric excess is 74%.

“Tautomers” are structurally distinct isomers that interconvert bytautomerization. “Tautomerization” is a form of isomerization andincludes prototropic or proton-shift tautomerization, which isconsidered a subset of acid-base chemistry. “Prototropictautomerization” or “proton-shift tautomerization” involves themigration of a proton accompanied by changes in bond order, often theinterchange of a single bond with an adjacent double bond. Wheretautomerization is possible (e.g., in solution), a chemical equilibriumof tautomers can be reached. An example of tautomerization is keto-enoltautomerization. A specific example of keto-enol tautomerization is theinterconversion of pentane-2,4-dione and 4-hydroxypent-3-en-2-onetautomers. Another example of tautomerization is phenol-ketotautomerization. A specific example of phenol-keto tautomerization isthe interconversion of pyridin-4-ol and pyridin-4(1H)-one tautomers.

“Protecting group” has the meaning conventionally associated with it inorganic synthesis, i.e. a group that selectively blocks one or morereactive sites in a multifunctional compound such that a chemicalreaction can be carried out selectively on another unprotected reactivesite and such that the group can readily be removed after the selectivereaction is complete. A variety of protecting groups are disclosed, forexample, in Wuts, Greene's Protective Groups in Organic Synthesis,5^(th) Ed., Wiley, New York, N.Y., 2014. For example, a hydroxyprotected form is where at least one of the hydroxy groups present in acompound is protected with a hydroxy protecting group. Likewise, aminesand other reactive groups may similarly be protected.

“Solvate” refers to a compound in physical association with one or moremolecules of a pharmaceutically acceptable solvent. It will beunderstood that the present chemical entities encompass the presentchemical entities and solvates of the compound, as well as mixturesthereof.

“Solvent,” “organic solvent,” and “inert solvent” each means a solventinert under the conditions of the reaction being described inconjunction therewith, including, for example, benzene, toluene,acetonitrile, tetrahydrofuran (“THF”), dimethylformamide (“DMF”),chloroform, methylene chloride (or dichloromethane), diethyl ether,methanol, N-methylpyrrolidone (“NMP”), pyridine and the like. Unlessspecified to the contrary, the solvents used in the reactions describedherein are inert organic solvents. Unless specified to the contrary, foreach gram of the limiting reagent, one cc (or mL) of solvent constitutesa volume equivalent.

Isolation and purification of the chemical entities and intermediatesdescribed herein can be effected, if desired, by any suitable separationor purification procedure such as, for example, filtration, extraction,crystallization, column chromatography, thin-layer chromatography orthick-layer chromatography, or a combination of these procedures.Specific illustrations of suitable separation and isolation procedurescan be had by reference to the examples hereinbelow. However, otherequivalent separation or isolation procedures can also be used.

When desired, the (R)- and (S)-isomers of the compounds of the presentinvention, if present, may be resolved by methods known to those skilledin the art, for example by formation of diastereoisomeric salts orcomplexes which may be separated, for example, by crystallization; viaformation of diastereoisomeric derivatives which may be separated, forexample, by crystallization, gas-liquid or liquid chromatography;selective reaction of one enantiomer with an enantiomer-specificreagent, for example enzymatic oxidation or reduction, followed byseparation of the modified and unmodified enantiomers; or gas-liquid orliquid chromatography in a chiral environment, for example on a chiralsupport, such as silica with a bound chiral ligand or in the presence ofa chiral solvent. Alternatively, a specific enantiomer may besynthesized by asymmetric synthesis using optically active reagents,substrates, catalysts or solvents, or by converting one enantiomer tothe other by asymmetric transformation.

The compounds described herein can be optionally contacted with apharmaceutically acceptable acid to form the corresponding acid additionsalts. Pharmaceutically acceptable forms of the compounds recited hereininclude pharmaceutically acceptable salts, chelates, non-covalentcomplexes, prodrugs, and mixtures thereof. In certain embodiments, thecompounds described herein are in the form of pharmaceuticallyacceptable salts. In addition, if the compound described herein isobtained as an acid addition salt, the free base can be obtained bybasifying a solution of the acid salt. Conversely, if the product is afree base, an addition salt, particularly a pharmaceutically acceptableaddition salt, may be produced by dissolving the free base in a suitableorganic solvent and treating the solution with an acid, in accordancewith conventional procedures for preparing acid addition salts from basecompounds. Those skilled in the art will recognize various syntheticmethodologies that may be used to prepare non-toxic pharmaceuticallyacceptable addition salts.

When ranges are used herein for physical properties, such as molecularweight, or chemical properties, such as chemical formulae, allcombinations and subcombinations of ranges and specific embodimentstherein are intended to be included. The term “about” when referring toa number or a numerical range means that the number or numerical rangereferred to is an approximation within experimental variability (orwithin statistical experimental error), and thus the number or numericalrange may vary from, for example, between 1% and 15% of the statednumber or numerical range.

Abbreviations used herein have their conventional meaning within thechemical and biological arts.

The following abbreviations and terms have the indicated meaningsthroughout:

DAST=Diethylaminosulfur trifluoride

DCM=Dichloromethane

MTBE=Methyl t-butyl ether

HATU=O-(7-azabenzotriazol-1-yl)-N,N,N′,N′-tetramethyluroniumhexafluorophosphate

NBS=N-Bromosuccinimide

NMP=N-Methyl-2-pyrrolidone

e.e. or ee=Enantiomeric excess

PPTS=Pyridinium p-toluenesulfonate

TLC=Thin Layer Chromatography

DMAP=4-Dimethylaminopyridine

DMF=N,N-Dimethylformamide

When stereochemistry is not specified, certain small molecules describedherein include, but are not limited to, when possible, their isomers,such as enantiomers and diastereomers, mixtures of enantiomers,including racemates, mixtures of diastereomers, and other mixturesthereof, to the extent they can be made by one of ordinary skill in theart by routine experimentation. In those situations, the singleenantiomers or diastereomers, i.e., optically active forms, can beobtained by asymmetric synthesis or by resolution of the racemates ormixtures of diastereomers. Resolution of the racemates or mixtures ofdiastereomers, if possible, can be accomplished, for example, byconventional methods such as crystallization in the presence of aresolving agent, or chromatography, using, for example, a chiralhigh-pressure liquid chromatography (HPLC) column. Furthermore, amixture of two enantiomers enriched in one of the two can be purified toprovide further optically enriched form of the major enantiomer byrecrystallization and/or trituration. In addition, such certain smallmolecules include Z- and E-forms (or cis- and trans-forms) of certainsmall molecules with carbon-carbon double bonds or carbon-nitrogendouble bonds. Where certain small molecules described herein exist invarious tautomeric forms, the term “certain small molecule” is intendedto include all tautomeric forms of the certain small molecule.

When “

” is drawn across a bond, it denotes where a bond disconnection orattachment occurs. For example, in the chemical structure shown below,

R^(a) is attached to the para position of a fluorophenyl ring through asingle bond. When R^(a) is phenyl, it can also be drawn as

The waved line “

” means a bond with undefined stereochemistry. For example,

represents a mixture of

When a bond is drawn across a ring, it means substitution at anon-specific ring atom or position. For example, in the structure shownbelow,

R^(b) may be attached to any one of the —CH₂— in the five-membered ring.

When a bold bond “

” appears two or more times in the same chemical structure, a mixture ofthe two cis isomers of the compound is described. For example,

represents a mixture of the two isomers

In one aspect, the present invention provides a method for treating aproliferative disorder in a subject in need thereof, comprisingadministering to said subject a HIF-2α inhibitor and animmunotherapeutic agent. In some embodiments, the HIF-2α inhibitor andthe immunotherapeutic agent are administered sequentially orsimultaneously. In some embodiments, the HIF-2α inhibitor and theimmunotherapeutic agent are more effective in treating the proliferativedisorder than either agent alone. In some embodiments, the HIF-2αinhibitor and the immunotherapeutic agent yield a synergistic effect intreating the proliferative disorder. The synergistic effect may be atherapeutic effect that is greater than either agent used alone incomparable amounts under comparable conditions. The synergistic effectmay be a therapeutic effect that is greater than results expected byadding the effects of each agent alone. In some embodiments, theproliferative disorder is a cancer condition. In some furtherembodiments, said cancer condition is a cancer selected from the groupconsisting of lung cancer, head and neck squamous cell carcinoma,pancreatic cancer, breast cancer, ovarian cancer, renal cell carcinoma,prostate cancer, neuroendocrine cancer, gastric cancer, bladder cancerand colon cancer. In another embodiment, the cancer condition is renalcell carcinoma. In still another embodiment, the cancer condition is acancer selected from the group consisting of non-small-cell lungcarcinoma, melanoma, renal cell carcinoma, colorectal cancer,castration-resistant prostate cancer, hepatocellular carcinoma, squamouscell carcinoma of the head and neck, carcinomas of the esophagus, ovary,gastrointestinal tract and breast, and a hematologic malignancy.

In a further embodiment, the present invention provides a method oftreating a cancer condition, wherein the HIF-2α inhibitor and theimmunotherapeutic agent together are effective in one or more ofinhibiting proliferation of cancer cells, inhibiting metastasis ofcancer cells, killing cancer cells, modulating an immune response andreducing severity or incidence of symptoms associated with the presenceof cancer cells. In some other embodiments, said method comprisesadministering to the cancer cells simultaneously or sequentially atherapeutically effective amount of a combination of a HIF-2α inhibitorand an immunotherapeutic agent. In some embodiments, the administrationtakes place in vitro. In other embodiments, the administration takesplace in vivo.

In some embodiments, the present invention provides a method of treatinga von Hippel-Lindau (VHL) disease, comprising administering to a subjectin need thereof a HIF-2α inhibitor and an immunotherapeutic agent. VHLdisease is an autosomal dominant syndrome that not only predisposespatients to kidney cancer (˜70% lifetime risk), but also tohemangioblastomas, pheochromocytoma and pancreatic neuroendocrinetumors. VHL disease results in tumors with constitutively active HIF-αproteins with the majority of these dependent on HIF-2α activity (Maher,et al. Eur. J. Hum. Genet. 19: 617-623, 2011). HIF-2α has been linked tocancers of the retina, adrenal gland and pancreas through both VHLdisease and activating mutations. Recently, gain-of-function HIF-2αmutations have been identified in erythrocytosis and paraganglioma withpolycythemia (Zhuang, et al. NEJM 367: 922-930, 2012; Percy, et al. NEJM358: 162-168, 2008; and Percy, et al. Am. J. Hematol. 87: 439-442,2012). Notably, a number of known HIF-2α target gene products (e.g.,VEGF, PDGF, and cyclin D1) have been shown to play pivotal roles incancers derived from kidney, liver, colon, lung, and brain. In fact,therapies targeted against one of the key HIF-2α regulated geneproducts, VEGF, have been approved for the treatment of these cancers.

In practicing any of the discussed methods, a HIF-2α inhibitor and animmunotherapeutic agent can be administered sequentially, wherein thetwo agents are introduced into a subject at two different time points.The two time points can be separated by more than 2 hours, 1 or moredays, 1 or more weeks, 1 or more months, or according to anyintermittent regimen schedule disclosed herein.

In some embodiments, the HIF-2α inhibitor and the immunotherapeuticagent are administered simultaneously. The two agents may form part ofthe same composition, or the two agents may be provided in one or moreunit doses.

In some other embodiments, the HIF-2α inhibitor or the immunotherapeuticagent are administered parenterally, orally, intraperitoneally,intravenously, intraarterially, transdermally, intramuscularly,liposomally, via local delivery by catheter or stent, subcutaneously,intraadiposally, or intrathecally.

As used herein, a therapeutically effective amount of a combination of aHIF-2α inhibitor and an immunotherapeutic agent refers to a combinationof a HIF-2α inhibitor and an immunotherapeutic agent, wherein thecombination is sufficient to effect the intended application, includingbut not limited to, disease treatment, as defined herein. Alsocontemplated in the subject methods is the use of a sub-therapeuticamount of a HIF-2α inhibitor and an immunotherapeutic agent incombination for treating an intended disease condition. The individualcomponents of the combination, though present in sub-therapeuticamounts, synergistically yield an efficacious effect and/or reduced aside effect in an intended application.

The amount of the HIF-2α inhibitor and the immunotherapeutic agentadministered may vary depending upon the intended application (in vitroor in vivo), or the subject and disease condition being treated, e.g.,the weight and age of the subject, the severity of the diseasecondition, the manner of administration and the like, which can readilybe determined by one of ordinary skill in the art.

Measuring an immune response and/or the inhibition of biological effectsof HIF-2α can comprise performing an assay on a biological sample, suchas a sample from a subject. Any of a variety of samples may be selected,depending on the assay. Examples of samples include, but are not limitedto blood samples (e.g. blood plasma or serum), exhaled breath condensatesamples, bronchioalveolar lavage fluid, sputum samples, urine samples,and tissue samples.

A subject being treated with a HIF-2α inhibitor and an immunotherapeuticagent may be monitored to determine the effectiveness of treatment, andthe treatment regimen may be adjusted based on the subject'sphysiological response to treatment. For example, if inhibition of abiological effect of HIF-2α inhibition is above or below a threshold,the dosing amount or frequency may be decreased or increased,respectively. Alternatively, the treatment regimen may be adjusted withrespect to an immune response. The methods can further comprisecontinuing the therapy if the therapy is determined to be efficacious.The methods can comprise maintaining, tapering, reducing, or stoppingthe administered amount of a compound or compounds in the therapy if thetherapy is determined to be efficacious. The methods can compriseincreasing the administered amount of a compound or compounds in thetherapy if it is determined not to be efficacious. Alternatively, themethods can comprise stopping therapy if it is determined not to beefficacious. In some embodiments, treatment with a HIF-2α inhibitor andan immunotherapeutic agent is discontinued if inhibition of thebiological effect is above or below a threshold, such as in a lack ofresponse or an adverse reaction. The biological effect may be a changein any of a variety of physiological indicators.

In one embodiment, an immunotherapeutic agent may comprise a PD-1inhibitor. In another embodiment, an immunotherapeutic agent maycomprise a CTLA-4 inhibitor. In still another embodiment, animmunotherapeutic agent may comprise a B7 inhibitor.

Exemplary PD-1 Inhibitors:

A PD-1 inhibitor suitable for use in the subject methods can be selectedfrom a variety of types of molecules. For example, the PD-1 inhibitorcan be a biological or chemical compound, such as an organic orinorganic molecule, peptide, peptide mimetic, antibody or anantigen-binding fragment of an antibody. Some exemplary classes ofagents suitable for use in the subject methods are detailed in thesections below. A PD-1 inhibitor for use in the present invention can beany PD-1 inhibitor that is known in the art, and can include any entitythat, upon administration to a patient, results in inhibition of thePD-1 pathway in the patient. A PD-1 inhibitor can inhibit PD-1 by anybiochemical mechanism, including disruption of any one or more ofPD-1/PD-L1, PD-1/PD-L2 and PD-L1/CD80 interactions.

In some embodiments, the PD-1 inhibitor is a molecule that inhibits thebinding of PD-1 to its ligand binding partners. In a specific aspect,the PD-1 ligand binding partners are PD-L1 and/or PD-L2. In anotherembodiment, a PD-1 inhibitor is a molecule that inhibits the binding ofPD-L1 to its binding partners. In a specific aspect, PD-L1 bindingpartners are PD-1 and/or CD80. In another embodiment, the PD-1 inhibitoris a molecule that inhibits the binding of PD-L2 to its bindingpartners. In a specific aspect, a PD-L2 binding partner is PD-1. Theinhibitor may be an antibody, an antigen binding fragment thereof, animmunoadhesin, a fusion protein or oligopeptide.

In some embodiments, the PD-1 inhibitor is an anti-PD-1 antibody. Insome further embodiments, the anti-PD-1 antibody is capable ofinhibiting binding between PD-1 and PD-L1. In another embodiment, theanti-PD-1 antibody is capable of inhibiting binding between PD-1 andPD-L2. In some embodiments, the PD-1 inhibitor is an anti-PD-L1antibody. In some embodiments, the anti-PD-L1 antibody is capable ofinhibiting binding between PD-L1 and PD-1 and/or between PD-L1 and CD80.In some embodiments, the PD-1 inhibitor is an anti-PD-L2 antibody. Insome further embodiments, the anti-PD-L2 antibody is capable ofinhibiting binding between PD-1 and PD-L2. In yet another embodiment,the PD-1 inhibitor is nivolumab or pembrolizumab.

Inhibition of the PD-1 pathway can enhance the immune response tocancerous cells in a patient. The interaction between PD-1 and PD-L1impairs T cell response as manifested by a decrease intumor-infiltrating lymphocytes (TILs) and a decrease in T-cell receptormediated proliferation, resulting in T cell energy, exhaustion orapoptosis, and immune evasion by the cancerous cells. This immunesuppression can be reversed by inhibiting the local interaction betweenPD-L1 and PD-1 using a PD-1 inhibitor, including, for example, ananti-PD-1 and/or an anti-PD-L1 Ab. A PD-1 inhibitor may improve orrestore antitumor T-cell functions.

Anti-PD-1 antibodies suitable for use in the invention can be generatedusing methods well known in the art. Exemplary PD-1 inhibitors include,but are not limited to: nivolumab (BMS936558), pembrolizumab (MK-3475),pidilizumab (CT-011), AMP-224, AMP-514, BMS-936559, RG7446 (MPDL3280A),MDX-1106 (Medarex Inc.), MSB0010718C, MEDI4736, and HenGrui mAB005 (WO15/085847). Further PD-1 antibodies and other PD-1 inhibitors includethose described in WO 04/056875, WO 06/121168, WO 07/005874, WO08/156712, WO 09/014708, WO 09/114335, WO 09/101611, WO 10/036959, WO10/089411, WO 10/027827, WO 10/077634, WO 11/066342, WO 12/145493, WO13/019906, WO 13/181452, WO 14/022758, WO 14/100079, WO 14/206107, WO15/036394, WO 15/085847, WO 15/112900, WO 15/112805, WO 15/112800, WO15/109124, WO 15/061668, WO 15/048520, WO 15/044900, WO 15/036927, WO15/035606; U.S. Pub. No. 2015/0071910; and U.S. Pat. Nos. 7,488,802;7,521,051; 7,595,048; 7,722,868; 7,794,710; 8,008,449; 8,354,509;8,383,796; 8,652,465; and 8,735,553; all of which are incorporatedherein by reference. Some anti-PD-1 antibodies are commerciallyavailable, for example from ABCAM® (AB 137132), BIOLEGEND® (EH12.2H7,RMP1-14) and AFFYMETRIX EBIOSCIENCE (J105, J116, M1H4).

Exemplary CTLA-4 Inhibitors:

A CTLA-4 inhibitor suitable for use in the subject methods can beselected from a variety of types of molecules. For example, the CTLA-4inhibitor can be a biological or chemical compound, such as an organicor inorganic molecule, peptide, peptide mimetic, antibody or anantigen-binding fragment of an antibody. Some exemplary classes ofagents suitable for use in the subject methods are detailed in thesections below. A CTLA-4 inhibitor for use in the present invention canbe any CTLA-4 inhibitor that is known in the art, and can include anyentity that, upon administration to a patient, results in inhibition ofthe CTLA-4 pathway in the patient. A CTLA-4 inhibitor can inhibit CTLA-4by any biochemical mechanism, including disruption of either one or bothof CTLA-4/CD80 and CTLA-4/CD86 interactions.

In some embodiments, the CTLA-4 inhibitor is a molecule that inhibitsthe binding of CTLA-4 to its ligand binding partners. In a specificaspect, the CTLA-4 ligand binding partners are CD80 and/or CD86. Inanother embodiment, a CTLA-4 inhibitor is a molecule that inhibits thebinding of CD80 to its binding partners. In a specific aspect, a CD80binding partner is CTLA-4. In another embodiment, the CTLA-4 inhibitoris a molecule that inhibits the binding of CD86 to its binding partners.In a specific aspect, a CD86 binding partner is CTLA-4. The inhibitormay be an antibody, an antigen binding fragment thereof, animmunoadhesin, a fusion protein or oligopeptide.

In some embodiments, the CTLA-4 inhibitor is an anti-CTLA-4 antibody. Insome further embodiments, the anti-CTLA-4 antibody is capable ofinhibiting binding between CTLA-4 and CD80. In another embodiment, theanti-CTLA-4 antibody is capable of inhibiting binding between CTLA-4 andCD86. In some embodiments, the CTLA-4 inhibitor is an anti-CD80antibody. In some embodiments, the anti-CD80 antibody is capable ofinhibiting binding between CTLA-4 and CD80. In some embodiments, theCTLA-4 inhibitor is an anti-CD86 antibody. In some further embodiments,the anti-CD86 antibody is capable of inhibiting binding between CTLA-4and CD86. In yet another embodiment, the CTLA-4 inhibitor istremelimumab or ipilimumab.

Inhibition of the CTLA-4 pathway can enhance the immune response tocancerous cells in a patient. The interaction between CTLA-4 and one ofits natural ligands, CD80 and CD86, delivers a negative regulatorysignal to T cells. This immune suppression can be reversed by inhibitingthe local interaction between CD80 or CD86 and CTLA-4 using a CTLA-4inhibitor, including, for example, an anti-CTLA-4 Ab, anti-CD80 Ab or ananti-CD86 Ab. A CTLA-4 inhibitor may improve or restore antitumor T-cellfunctions.

Anti-CTLA-4 antibodies suitable for use in the invention can begenerated using methods well known in the art. Exemplary CTLA-4inhibitors include but are not limited to tremelimumab and ipilimumab(also known as 10D1 or MDX-010). Further CTLA-4 antibodies and otherCTLA-4 inhibitors include those described in WO 98/042752, WO 00/037504,WO 01/014424 and WO 04/035607; U.S. Pub. Nos. 2002/0039581, 2002/086014and 2005/0201994; U.S. Pat. Nos. 5,811,097; 5,855,887; 5,977,318;6,051,227; 6,207,156; 6,682,736; 6,984,720; 7,109,003; 7,132,281;7,605,238; 8,143,379; 8,318,916; 8,435,516; 8,784,815; and 8,883,984; EPPat. No. 1212422; Hurwitz et al., PNAS 1998, 95(17): 10067-10071;Camacho et al., J Clin Oncology 2004, 22(145): abstract no. 2505(antibody CP-675206); and Mokyr, et al., Cancer Research 1998,58:5301-5304; all of which are incorporated herein by reference.

Exemplary HIF-2α Inhibitors:

A HIF-2α inhibitor suitable for use in the subject methods can beselected from a variety of types of molecules. For example, the HIF-2αinhibitor can be a biological or chemical compound, such as a simple orcomplex organic or inorganic molecule, peptide, peptide mimetic, protein(e.g. antibody), liposome, or a polynucleotide (e.g. small interferingRNA, microRNA, anti-sense, aptamer, ribozyme, or triple helix). Someexemplary classes of chemical compounds suitable for use in the subjectmethods are detailed in the sections below. A HIF-2α inhibitor for usein the present invention can be any HIF-2α inhibitor that is known inthe art, and can include any chemical entity that, upon administrationto a patient, results in inhibition of HIF-2α in the patient.

In general, a HIF-2α inhibitor is a compound that inhibits one or morebiological effects of HIF-2α. Examples of biological effects of HIF-2αinclude, but are not limited to, heterodimerization of HIF-2α to HIF-1β,HIF-2α target gene expression, VEGF gene expression, and VEGF proteinsecretion. In some embodiments, the HIF-2α inhibitor is selective forHIF-2α, such that the inhibitor inhibits heterodimerization of HIF-2α toHIF-1β but not heterodimerization of HIF-1α to HIF-13. Such biologicaleffects may be inhibited by about or more than about 10%, 20%, 30%, 40%,50%, 60%, 70%, 80%, 90%, 95%, or more.

Hypoxia-inducible factors (HIFs), like HIF-2α, are transcription factorsthat respond to changes in available oxygen in the cellular environment(e.g. a decrease in oxygen, or hypoxia). The HIF signaling cascademediates the effects of hypoxia, the state of low oxygen concentration,on the cell. Hypoxia often keeps cells from differentiating. However,hypoxia promotes the formation of blood vessels, and is important forthe formation of a vascular system in embryos, and cancer tumors. Thehypoxia in wounds also promotes the migration of keratinocytes and therestoration of the epithelium. A HIF-2α inhibitor of the presentdisclosure may be administered in an amount effective in reducing anyone or more of such effects of HIF-2α activity.

HIF-2α activity can be inhibited by inhibiting heterodimerization ofHIF-2α to HIF-1β (ARNT), such as with inhibitor compounds disclosedherein. A variety of methods for measuring HIF-2α dimerization areavailable. In some embodiments, the HIF-2α inhibitor binds the PAS-Bdomain cavity of HIF-2α.

Inhibition of heterodimerization of HIF-2α to HIF-1β (ARNT) may also bedetermined by a reduction in HIF-2α target gene mRNA expression. mRNAquantitation can be performed using real-time PCR technology. (Wong, etal, “Real-time PCR for mRNA quantitation”, 2005. BioTechniques 39, 1:1-1.). Yet another method for determining inhibition ofheterodimerization of HIF-2α to HIF1β (ARNT) is byco-immunoprecipitation.

As described herein, HIF-2α is a transcription factor that playsimportant roles in regulating expression of target genes. Non-limitingexamples of HIF-2α target gene include HMOX1, SFTPA1, CXCR4, PAI1, BDNF,hTERT, ATP7A, and VEGF. For instance, HIF-2α is an activator of VEGF.Further non-limiting examples of HIF-2α target genes include HMOX1, EPO,CXCR4, PAI1, CCND1, CLUT1, IL6, and VEGF. A HIF-2α inhibitor of thepresent disclosure may be administered in an amount effective inreducing expression of any one or more of genes induced by HIF-2αactivity. A variety of methods is available for the detection of geneexpression levels, and includes the detection of gene transcriptionproducts (polynucleotides) and translation products (polypeptides). Forexample, gene expression can be detected and quantified at the DNA, RNAor mRNA level. Various methods that have been used to quantify mRNAinclude in situ hybridization techniques, fluorescent in situhybridization techniques, reporter genes, RNase protection assays,Northern blotting, reverse transcription (RT)-PCR, SAGE, DNA microarray,tiling array, and RNA-seq. Examples of methods for the detection ofpolynucleotides include, but are not limited to selective colorimetricdetection of polynucleotides based on the distance-dependent opticalproperties of gold nanoparticles, and solution phase detection ofpolynucleotides using interacting fluorescent labels and competitivehybridization. Examples for the detection of proteins include, but arenot limited to microscopy and protein immunostaining, proteinimmunoprecipitation, immunoelectrophoresis, western blot, BCA assay,spectrophotometry, mass spectrophotometry and enzyme assay.

In some embodiments, inhibition of HIF-2α is characterized by a decreasein VEGF gene expression. The decrease may be measured by any of avariety of methods, such as those described herein. As a furtherexample, the mRNA expression level of VEGF can be measured byquantitative PCR (QT-PCR), microarray, RNA-seq and nanostring. Asanother example, an ELISA assay can be used to measure the level VEGFprotein secretion.

In one aspect, the invention provides a compound which is an inhibitorof HIF-2α of Formula I:

or a pharmaceutically acceptable salt or prodrug thereof, wherein:

X is CR⁵ or N;

Y is CR⁶ or N;

Z is —O—, —S—, —C(HR⁷)—, —N(R⁸)— or absent;

R¹ is alkyl, heteroalkyl, cycloalkyl, heterocycloalkyl, aryl,heteroaryl, acyl or cyano;

R² is nitro, carboxaldehyde, carboxyl, ester, amido, cyano, halo,sulfonyl, alkyl, alkenyl, alkynyl or heteroalkyl;

R³ is hydrogen, halo, cyano, alkyl, heteroalkyl, alkenyl, alkynyl,amino, carboxaldehyde, carboxylic acid, oxime, ester, amido or acyl; orR² and R³ taken together form a cyclic moiety;

R⁴ is nitro, halo, cyano, alkyl, cycloalkyl, heteroaryl, carboxyl,sulfinyl, sulfonamidyl, sulfonyl or sulfoximinyl; and

R⁵, R⁶, R⁷ and R⁸ are independently hydrogen, halo, hydroxy, cyano,alkyl or alkoxy.

In one aspect, the invention provides a compound which is an inhibitorof HIF-2α of Formula I:

or a pharmaceutically acceptable salt or prodrug thereof, wherein:

X is CR⁵ or N;

Y is CR⁶ or N;

Z is —O—, —S—, —S(O)—, —S(O)₂—, —C(O)—, —C(HR⁷)—, —N(R⁸)—, C₁-C₃alkylene, C₁-C₃ heteroalkylene, C₁-C₃ alkenylene or absent;

R¹ is alkyl, heteroalkyl, cycloalkyl, heterocycloalkyl, aryl,heteroaryl, acyl or cyano;

R² is nitro, carboxaldehyde, carboxyl, ester, amido, cyano, halo,sulfonyl, alkyl, alkenyl, alkynyl or heteroalkyl;

R³ is hydrogen, halo, cyano, alkyl, heteroalkyl, alkenyl, alkynyl,amino, carboxaldehyde, carboxylic acid, oxime, ester, amido or acyl; orR² and R³ taken together form a cyclic moiety;

R⁴ is nitro, halo, cyano, alkyl, cycloalkyl, heteroaryl, carboxyl,sulfinyl, sulfonamidyl, sulfonyl or sulfoximinyl; and

R⁵, R⁶, R⁷ and R⁸ are independently hydrogen, halo, hydroxy, cyano,alkyl or alkoxy.

In some embodiments, for a compound of Formula I, R¹ is further selectedfrom alkyl, heteroalkyl, cycloalkyl, cycloalkenyl, heterocycloalkyl,aryl, heteroaryl, acyl or cyano.

In some embodiments, R¹ is phenyl or monocyclic heteroaryl. In somefurther embodiments, R¹ is phenyl or pyridyl, optionally substitutedwith one or more substituents selected from the group consisting ofhalo, alkyl, alkoxy and cyano. In a further embodiment, thesubstituent(s) is selected from the group consisting of halo, C₁-C₄alkyl, C₁-C₄ alkoxy and cyano.

In some embodiments, R¹ is bicyclic heteroaryl. In a further embodiment,the bicyclic heteroaryl is substituted with one or more substituentsselected from the group consisting of halo, C₁-C₄ alkyl, C₁-C₄ alkoxyand cyano.

In some embodiments, R¹ is pyridyl N-oxide. In a further embodiment, thepyridyl N-oxide is substituted with one or more substituents selectedfrom the group consisting of halo, C₁-C₄ alkyl, C₁-C₄ alkoxy and cyano.

In some embodiments, R¹ is

wherein the aryl ring may optionally be substituted with one or moresubstituents selected from the group consisting of cyano, halo, alkyland alkoxy. In a further embodiment, the substituent(s) is selected fromthe group consisting of halo, C₁-C₄ alkyl, C₁-C₄ alkoxy and cyano.

In some embodiments, R¹ is

wherein W¹ is N or CR¹⁰, R⁹ is cyano, halo, alkyl or alkoxy, and R¹⁰ ishydrogen, cyano, halo, alkyl or alkoxy. In a further embodiment, R⁹ iscyano, halo, C₁-C₄ alkyl or C₁-C₄ alkoxy, and R¹⁰ is hydrogen, cyano,halo, C₁-C₄ alkyl or C₁-C₄ alkoxy.

In some embodiments, R¹ is selected from the group consisting of:

and the rings specified for R¹ may optionally be substituted with one ormore substituents described for aryl and heteroaryl. In a furtherembodiment, the substituent(s) is selected from the group consisting ofhalo, C₁-C₄ alkyl, C₁-C₄ alkoxy and cyano.

In some embodiments, R¹ is selected from the group consisting of:

and the rings specified for R¹ may optionally be substituted with one ormore substituents described for cycloalkyl. In a further embodiment, thesubstituent(s) is selected from the group consisting of halo, hydroxy,C₁-C₄ alkyl, C₁-C₄ alkoxy, cyano and oxo.

In some embodiments, R¹ is cycloalkyl. In other embodiments, R¹ isheterocycloalkyl. In a further embodiment, R¹ is C₃-C₆ cycloalkyl orC₃-C₆ heterocycloalkyl. In yet a further embodiment, R¹ is cyclobutyl.In some embodiments, said cycloalkyl, cyclobutyl or heterocycloalkyl mayoptionally be substituted with one or more substituents described forcycloalkyl or heterocycloalkyl. In a further embodiment, thesubstituent(s) is selected from the group consisting of halo, C₁-C₄alkyl, C₁-C₄ alkoxy and cyano. In another further embodiment, thesubstituent(s) is at least one fluoro.

In some embodiments, R¹ is acyl or cyano. In a further embodiment, R¹ isacetyl.

In some embodiments, R¹ is alkyl. In a further embodiment, the alkyl issubstituted with at least one substituent(s) selected from the groupconsisting of halo, C₁-C₄ alkyl, C₁-C₄ alkoxy and cyano. In anotherfurther embodiment, the alkyl is substituted with at least one fluoro.In some embodiments, R¹ is heteroalkyl.

In some embodiments, R¹ is selected from the group consisting of:

wherein each of the members may optionally be substituted with one ormore substituents selected from the group consisting of cyano, halo,alkyl and alkoxy. In a further embodiment, the substituent(s) isselected from the group consisting of fluoro, C₁-C₄ alkyl, C₁-C₄ alkoxyand cyano.

In some embodiments, R² is cyano, halo or alkyl. In some embodiments, R²is halo or alkyl. In some embodiments, R² is fluoro, chloro, bromo oriodo. In some embodiments, R² is fluoroalkyl. In some furtherembodiments, R² is —CH₂F, —CHF₂ or —CF₃. In another embodiment, R² ishydrogen. In some other embodiments, R² is heteroalkyl, alkenyl oralkynyl.

In some embodiments, R³ is hydrogen, halo, cyano, alkyl, alkenyl,heteroalkyl or acyl; or R² and R³ taken together form a cyclic moiety.In a further embodiment, R³ is halo, cyano or alkyl. In yet a furtherembodiment, R³ is —(CH₂)_(n)OH, wherein n is 1, 2 or 3. In still afurther embodiment, R³ is —CH₂OH.

In some embodiments, R² and R³ taken together with the atoms to whichthey are attached form a 5- or 6-membered carbocycle with at least onesp³ hybridized carbon. Representative compounds with the carbocycleinclude, but are not limited to, the following:

wherein the carbocycle formed by linking R² and R³ may be optionallysubstituted with fluoro, chloro, hydroxy, alkyl or heteroalkyl. In afurther embodiment, the substituent(s) is selected from the groupconsisting of halo, C₁-C₄ alkyl, C₁-C₄ alkoxy and cyano. In yet otherembodiments, the substituent(s) is cycloalkyl or heterocycloalkyl andshares one or more ring atoms with the carbocycle formed by linking R²and R³. In some embodiments, the substituent(s) is C₃-C₅ cycloalkyl orC₃-C₅ heterocycloalkyl. In other embodiments, the substituent is oxo.

In some embodiments, R² and R³ taken together with the atoms to whichthey are attached form a 5- or 6-membered heterocycle, including, butnot limited to, a lactone or lactol, wherein said heterocycle may beoptionally substituted with fluoro, chloro, hydroxy, alkyl orheteroalkyl. In a further embodiment, the substituent(s) is selectedfrom the group consisting of halo, C₁-C₄ alkyl, C₁-C₄ alkoxy and cyano.

In some embodiments, R⁴ is halo, cyano, fluoroalkyl, sulfinyl,sulfonamidyl, sulfonyl, sulfoximinyl or fluoroalkylsulfonyl. In someembodiments, R⁴ is cyano, fluoroalkyl, sulfonamidyl, sulfinyl, sulfonylor sulfoximinyl. In some embodiments, R⁴ is fluoroalkyl, sulfonamidyl,sulfonyl, sulfoximinyl or fluoroalkylsulfonyl. In a further embodiment,R⁴ is fluoroalkyl. In yet another embodiment, R⁴ is sulfonyl. In stillanother embodiment, R⁴ is alkylsulfonyl.

In some embodiments, R⁴ is —S(═O)₂R^(a), wherein R^(a) is alkyl orcycloalkyl. In a further embodiment, R^(a) is C₁-C₄ alkyl, optionallysubstituted with one or more fluorines. Suitable examples offluorine-substituted C₁-C₄ alkyl include, but are not limited to, —CH₂F,—CHF₂, —CF₃, —CH₂CF₃, —CH₂CHF₂, —CH₂CH₂F, —CHFCH₃ and —CF₂CH₃. In stilla further embodiment, R^(a) is methyl, optionally substituted with oneor more fluorines.

In some embodiments, R⁴ is —S(═O)(═NR^(b))R^(a), wherein R^(a) is alkylor cycloalkyl and R^(b) is hydrogen, cyano or alkyl. In a furtherembodiment, R^(a) is C₁-C₄ alkyl, optionally substituted with one ormore fluorines. Suitable examples of fluorine-substituted C₁-C₄ alkylinclude, but are not limited to, —CH₂F, —CHF₂, —CF₃, —CH₂CF₃, —CH₂CHF₂,—CH₂CH₂F, —CHFCH₃ and —CF₂CH₃.

In some embodiments, R⁴ is —S(═O)₂N(R^(a))₂, wherein each R^(a) isindependently hydrogen, alkyl, heteroalkyl, cycloalkyl orheterocycloalkyl, and at least one R^(a) is hydrogen. In a furtherembodiment, both R^(a)s are hydrogen. In another further embodiment, oneR^(a) is hydrogen and the other R^(a) is C₁-C₄ alkyl.

In some embodiments, R⁴ is selected from the group consisting of —CN,—CF₃, —S(═O)CH₃, —S(═O)₂CH₃, —S(═O)₂CH₂F, —S(═O)₂CHF₂, —S(═O)₂CF₃,—S(═O)₂NH₂, —S(═O)₂NHCH₃, —S(═O)(═NH)CH₃, —S(═O)(═NH)CH₂F,—S(═O)(═NH)CHF₂, —S(═O)(═NH)CF₃, —S(═O)(═N—CN)CH₃, —S(═O)(═N—CN)CH₂F,—S(═O)(═N—CN)CHF₂ and —S(═O)(═N—CN)CF₃.

In some embodiments, R⁵ is hydrogen. In some other embodiments, R⁵ isC₁-C₄ alkyl or C₁-C₄ alkoxy. In a further embodiment, R⁵ is methyl.

In some embodiments, R⁶ is hydrogen. In some other embodiments, R⁶ isC₁-C₄ alkyl or C₁-C₄ alkoxy. In a further embodiment, R⁶ is methyl.

In some embodiments, R⁷ is hydrogen. In some other embodiments, R⁷ isC₁-C₄ alkyl or C₁-C₄ alkoxy. In a further embodiment, R⁷ is methyl.

In some embodiments, R⁸ is hydrogen. In some other embodiments, R⁸ isC₁-C₄ alkyl or C₁-C₄ alkoxy. In a further embodiment, R⁸ is methyl.

In some embodiments, R³ is hydrogen, R⁴ is —S(═O)₂R^(a) or—S(═O)(═NR^(b))R^(c), wherein R^(a) is fluoroalkyl, R^(b) is hydrogen,cyano or alkyl and R^(c) is alkyl. In a further embodiment, R¹ isselected from the group consisting of

wherein W¹ is N or CR¹⁰, R⁹ is cyano, halo, alkyl or alkoxy, and R¹⁰ ishydrogen, cyano, halo, alkyl or alkoxy; and

may optionally be substituted with one or more substituents selectedfrom the group consisting of cyano, halo, alkyl and alkoxy. In a furtherembodiment, the alkyl is C₁-C₄ alkyl. In another further embodiment, thealkoxy is C₁-C₄ alkoxy.

In some embodiments, each of R² and R³ is independently alkyl and R⁴ iscyano, fluoroalkyl, sulfonamidyl, sulfinyl, sulfonyl, sulfoximinyl orfluoroalkylsulfonyl.

In some embodiments, R³ is —CH₂OH. In a further embodiment, R⁴ is cyano,fluoroalkyl, sulfonamidyl, sulfinyl, sulfonyl, sulfoximinyl orfluoroalkylsulfonyl and R⁵ is hydrogen. In still a further embodiment,R² is cyano, halo or alkyl.

In some embodiments, R¹ is phenyl or monocyclic heteroaryl; R² is nitro,halo, cyano or alkyl; R³ is halo, cyano or alkyl; R⁴ is cyano,fluoroalkyl, sulfonamidyl, sulfinyl, sulfonyl, sulfoximinyl orfluoroalkylsulfonyl. In a further embodiment, R⁴ is selected from thegroup consisting of —CN, —CF₃, —S(═O)CH₃, —S(═O)₂CH₃, —S(═O)₂CH₂F,—S(═O)₂CHF₂, —S(═O)₂CF₃, —S(═O)₂NH₂, —S(═O)₂NHCH₃, —S(═O)(═NH)CH₃,—S(═O)(═NH)CH₂F, —S(═O)(═NH)CHF₂, —S(═O)(═NH)CF₃, —S(═O)(═N—CN)CH₃,—S(═O)(═N—CN)CH₂F, —S(═O)(═N—CN)CHF₂ and —S(═O)(═N—CN)CF₃. In still afurther embodiment, R⁵ is hydrogen.

In some embodiments, R¹ is bicyclic heteroaryl; R² is nitro, halo, cyanoor alkyl; R³ is halo, cyano or alkyl; R⁴ is cyano, fluoroalkyl,sulfonamidyl, sulfinyl, sulfonyl, sulfoximinyl or fluoroalkylsulfonyl;and R⁵ is hydrogen.

In some embodiments, R¹ is phenyl, monocyclic heteroaryl or bicyclicheteroaryl; R² is halo, cyano or alkyl; R³ is halo, cyano or alkyl; R⁴is cyano, fluoroalkyl, sulfonamidyl, sulfinyl, sulfonyl, sulfoximinyl orfluoroalkylsulfonyl; and R⁵ is hydrogen.

In some embodiments, R² and R³ together with the atoms to which they areattached form a 5- or 6-membered carbocycle with at least one sp³hybridized carbon; R⁴ is cyano, fluoroalkyl, sulfonamidyl, sulfinyl,sulfonyl, sulfoximinyl or fluoroalkylsulfonyl; and R⁵ is hydrogen. In afurther embodiment, R¹ is phenyl or monocyclic heteroaryl. In anotherfurther embodiment, R¹ is bicyclic heteroaryl.

In some embodiments, X is N and Y is CR⁶. In other embodiments, X is CR⁵and Y is N. In still other embodiments, X is N and Y is N. In yet otherembodiments, X is CR⁵ and Y is CR⁶.

In some embodiments, Z is —O—, —S—, —S(O)—, —S(O)₂—, —S(O₂)N(R⁸)—,—C(O)—, —C(O)O—, —C(HR⁷)—, —N(R⁸)—, —C(O)N(R⁸)—, alkylene, alkenylene,alkynylene, heteroalkylene, or absent. In some embodiments, Z is —O—,—S—, —S(O)—, —S(O)₂—, —C(O)—, —C(HR⁷)—, —N(R⁸)—, C₁-C₃ alkylene, C₁-C₃heteroalkylene, C₁-C₃ alkenylene or absent. In some embodiments, Z is—O—. In other embodiments, Z is —S—. In further embodiments, Z is—C(HR⁷)—. In yet other embodiments, Z is —N(R⁸). In some embodiments, Zis absent.

In another aspect, the invention provides a compound of Formula I-A:

or a pharmaceutically acceptable salt or prodrug thereof, wherein:

X is CR⁵ or N;

Y is CR⁶ or N;

Z is —O—, —S—, —C(HR⁷)—, —N(R⁸)— or absent;

R² is nitro, carboxaldehyde, carboxylic acid, ester, amido, cyano, halo,sulfonyl or alkyl;

R³ is hydrogen, halo, cyano, alkyl, heteroalkyl, alkenyl, alkynyl,amino, oxime or acyl; or R² and R³ taken together form a cyclic moiety;

R⁴ is nitro, halo, cyano, alkyl, sulfinyl, sulfonamidyl, sulfonyl orsulfoximinyl; R⁵, R⁶, R⁷ and R⁸ are independently hydrogen, halo,hydroxy, cyano, alkyl or alkoxy;

W¹ is N or CR¹⁰;

R⁹ is cyano, halo, alkyl or alkoxy; and

R¹⁰ is hydrogen, cyano, halo, alkyl or alkoxy.

In another aspect, the invention provides a compound of Formula I-A:

or a pharmaceutically acceptable salt or prodrug thereof, wherein:

X is CR⁵ or N;

Y is CR⁶ or N;

Z is —O—, —S—, —S(O)—, —S(O)₂—, —C(O)—, —C(HR⁷)—, —N(R⁸)—, C₁-C₃alkylene, C₁-C₃ heteroalkylene, C₁-C₃ alkenylene or absent;

R² is nitro, carboxaldehyde, carboxylic acid, ester, amido, cyano, halo,sulfonyl or alkyl;

R³ is hydrogen, halo, cyano, alkyl, heteroalkyl, alkenyl, alkynyl,amino, oxime or acyl; or R² and R³ taken together form a cyclic moiety;

R⁴ is nitro, halo, cyano, alkyl, sulfinyl, sulfonamidyl, sulfonyl orsulfoximinyl;

R⁵, R⁶, R⁷ and R⁸ are independently hydrogen, halo, hydroxy, cyano,alkyl or alkoxy;

W¹ is N or CR¹⁰;

R⁹ is cyano, halo, alkyl or alkoxy; and

R¹⁰ is hydrogen, cyano, halo, alkyl or alkoxy.

In some embodiments, R² is cyano, halo or alkyl. In some embodiments, R²is halo or alkyl. In some embodiments, R² is fluoro, chloro, bromo oriodo. In some embodiments, R² is fluoroalkyl. In some furtherembodiments, R² is —CH₂F, —CHF₂ or —CF₃.

In some embodiments, R³ is hydrogen, halo, cyano, alkyl, alkenyl,heteroalkyl or acyl; or R² and R³ taken together form a cyclic moiety.In a further embodiment, R³ is halo, cyano or alkyl. In yet a furtherembodiment, R³ is —(CH₂)_(n)OH, wherein n is 1, 2 or 3. In still afurther embodiment, R³ is —CH₂OH.

In some embodiments, R² and R³ taken together with the atoms to whichthey are attached form a 5- or 6-membered carbocycle with at least onesp³ hybridized carbon. Representative compounds with the carbocycleinclude, but are not limited to, the following:

wherein the carbocycle formed by linking R² and R³ may be optionallysubstituted with fluoro, chloro, hydroxy, alkyl or heteroalkyl. In afurther embodiment, the substituent(s) is selected from the groupconsisting of halo, C₁-C₄ alkyl, C₁-C₄ alkoxy and cyano. In yet otherembodiments, the substituent(s) is cycloalkyl or heterocycloalkyl andshares one or more ring atoms with the carbocycle formed by linking R²and R³. In some embodiments, the substituent(s) is C₃-C₅ cycloalkyl orC₃-C₅ heterocycloalkyl. In other embodiments, the substituent is oxo.

In some embodiments, R² and R³ taken together with the atoms to whichthey are attached form a 5- or 6-membered heterocycle, including, butnot limited to, a lactone or lactol, wherein said heterocycle may beoptionally substituted with fluoro, chloro, hydroxy, alkyl orheteroalkyl. In a further embodiment, the substituent(s) is selectedfrom the group consisting of halo, C₁-C₄ alkyl, C₁-C₄ alkoxy and cyano.

In some embodiments, R⁴ is halo, cyano, fluoroalkyl, sulfinyl,sulfonamidyl, sulfonyl, sulfoximinyl or fluoroalkylsulfonyl. In someembodiments, R⁴ is cyano, fluoroalkyl, sulfonamidyl, sulfinyl, sulfonylor sulfoximinyl. In some embodiments, R⁴ is fluoroalkyl, sulfonamidyl,sulfonyl or sulfoximinyl. In a further embodiment, R⁴ is fluoroalkyl. Inyet another embodiment, R⁴ is sulfonyl. In still another embodiment, R⁴is alkylsulfonyl.

In some embodiments, R⁴ is —S(═O)₂R^(a), wherein R^(a) is alkyl orcycloalkyl. In a further embodiment, R^(a) is C₁-C₄ alkyl, optionallysubstituted with one or more fluorines. Suitable examples offluorine-substituted C₁-C₄ alkyl include, but are not limited to, —CH₂F,—CHF₂, —CF₃, —CH₂CF₃, —CH₂CHF₂, —CH₂CH₂F, —CHFCH₃ and —CF₂CH₃. In stilla further embodiment, R^(a) is methyl, optionally substituted with oneor more fluorines.

In some embodiments, R⁴ is —S(═O)(═NR^(b))R^(a), wherein R^(a) is alkylor cycloalkyl and R^(b) is hydrogen, cyano or alkyl. In a furtherembodiment, R^(a) is C₁-C₄ alkyl, optionally substituted with one ormore fluorines. Suitable examples of fluorine-substituted C₁-C₄ alkylinclude, but are not limited to, —CH₂F, —CHF₂, —CF₃, —CH₂CF₃, —CH₂CHF₂,—CH₂CH₂F, —CHFCH₃ and —CF₂CH₃.

In some embodiments, R⁴ is —S(═O)₂N(R^(a))₂, wherein each R^(a) isindependently hydrogen, alkyl, heteroalkyl, cycloalkyl orheterocycloalkyl, and at least one R^(a) is hydrogen. In a furtherembodiment, both R^(a)s are hydrogen. In another further embodiment, oneR^(a) is hydrogen and the other R^(a) is C₁-C₄ alkyl.

In some embodiments, R⁴ is selected from the group consisting of —CN,—CF₃, —S(═O)CH₃, —S(═O)₂CH₃, —S(═O)₂CH₂F, —S(═O)₂CHF₂, —S(═O)₂CF₃,—S(═O)₂NH₂, —S(═O)₂NHCH₃, —S(═O)(═NH)CH₃, —S(═O)(═NH)CH₂F,—S(═O)(═NH)CHF₂, —S(═O)(═NH)CF₃, —S(═O)(═N—CN)CH₃, —S(═O)(═N—CN)CH₂F,—S(═O)(═N—CN)CHF₂ and —S(═O)(═N—CN)CF₃.

In some embodiments, R⁵ is hydrogen. In some other embodiments, R⁵ isC₁-C₄ alkyl or C₁-C₄ alkoxy. In a further embodiment, R⁵ is methyl.

In some embodiments, R⁶ is hydrogen. In some other embodiments, R⁶ isC₁-C₄ alkyl or C₁-C₄ alkoxy. In a further embodiment, R⁶ is methyl.

In some embodiments, R⁷ is hydrogen. In some other embodiments, R⁷ isC₁-C₄ alkyl or C₁-C₄ alkoxy. In a further embodiment, R⁷ is methyl.

In some embodiments, R⁸ is hydrogen. In some other embodiments, R⁸ isC₁-C₄ alkyl or C₁-C₄ alkoxy. In a further embodiment, R⁸ is methyl.

In some embodiments, R⁹ is cyano, halo, C₁-C₄ alkyl or C₁-C₄ alkoxy.

In some embodiments, R¹⁰ is hydrogen, cyano, halo, C₁-C₄ alkyl or C₁-C₄alkoxy.

In some embodiments, R² and R³ taken together with the atoms to whichthey are attached form a 5- or 6-membered carbocycle with at least onesp³ hybridized carbon and R⁴ is cyano, fluoroalkyl, sulfonamidyl,sulfinyl, sulfonyl, sulfoximinyl or fluoroalkylsulfonyl.

In some embodiments, R³ is —CH₂OH and R⁴ is cyano, fluoroalkyl,sulfonamidyl, sulfonyl or sulfoximinyl. In a further embodiment, R⁵ ishydrogen. In still a further embodiment, R² is cyano, halo or alkyl.

In some embodiments, R² is halo, cyano or alkyl; R³ is CH₂OH; R⁴ iscyano, fluoroalkyl, sulfonamidyl, sulfonyl or sulfoximinyl. In a furtherembodiment, R⁴ is selected from the group consisting of —CN, —CF₃,—S(═O)CH₃, —S(═O)₂CH₃, —S(═O)₂CH₂F, —S(═O)₂CHF₂, —S(═O)₂CF₃, —S(═O)₂NH₂,—S(═O)₂NHCH₃, —S(═O)(═NH)CH₃, —S(═O)(═NH)CH₂F, —S(═O)(═NH)CHF₂,—S(═O)(═NH)CF₃, —S(═O)(═N—CN)CH₃, —S(═O)(═N—CN)CH₂F, —S(═O)(═N—CN)CHF₂and —S(═O)(═N—CN)CF₃.

In some embodiments, X is N and Y is CR⁶. In other embodiments, X is CR⁵and Y is N. In still other embodiments, X is N and Y is N. In yet otherembodiments, X is CR⁵ and Y is CR⁶.

In some embodiments, Z is —O—, —S—, —S(O)—, —S(O)₂—, —S(O₂)N(R⁸)—,—C(O)—, —C(O)O—, —C(HR⁷)—, —N(R⁸)—, —C(O)N(R⁸)—, alkylene, alkenylene,alkynylene, heteroalkylene, or absent. In some embodiments, Z is —O—,—S—, —S(O)—, —S(O)₂—, —C(O)—, —C(HR⁷)—, —N(R⁸)—, C₁-C₃ alkylene, C₁-C₃heteroalkylene, C₁-C₃ alkenylene or absent. In some embodiments, Z is—O—. In other embodiments, Z is —S—. In further embodiments, Z is—C(HR⁷)—. In yet other embodiments, Z is —N(R⁸). In some embodiments, Zis absent.

In another aspect, the invention provides a compound of Formula I-B:

or a pharmaceutically acceptable salt or prodrug thereof, wherein:

X is CR⁵ or N;

Y is CR⁶ or N;

Z is —O—, —S—, —C(HR⁷)—, —N(R⁸)— or absent;

R² is nitro, carboxaldehyde, carboxylic acid, ester, amido, cyano, halo,sulfonyl or alkyl;

R³ is hydrogen, halo, cyano, alkyl, heteroalkyl, alkenyl, alkynyl,amino, oxime or acyl; or R² and R³ taken together form a cyclic moiety;

R⁴ is nitro, halo, cyano, alkyl, sulfinyl, sulfonamidyl, sulfonyl orsulfoximinyl;

R⁵, R⁶, R⁷ and R⁸ are independently hydrogen, halo, hydroxy, cyano,alkyl or alkoxy;

R^(c) is hydrogen, cyano, halo, alkyl or alkoxy; and

n′ is 0, 1, 2, 3 or 4.

In another aspect, the invention provides a compound of Formula I-B:

or a pharmaceutically acceptable salt or prodrug thereof, wherein:

X is CR⁵ or N;

Y is CR⁶ or N;

Z is —O—, —S—, —S(O)—, —S(O)₂—, —C(O)—, —C(HR⁷)—, —N(R⁸)—, C₁-C₃alkylene, C₁-C₃ heteroalkylene, C₁-C₃ alkenylene or absent;

R² is nitro, carboxaldehyde, carboxylic acid, ester, amido, cyano, halo,sulfonyl or alkyl;

R³ is hydrogen, halo, cyano, alkyl, heteroalkyl, alkenyl, alkynyl,amino, oxime or acyl; or R² and R³ taken together form a cyclic moiety;

R⁴ is nitro, halo, cyano, alkyl, sulfinyl, sulfonamidyl, sulfonyl orsulfoximinyl;

R⁵, R⁶, R⁷ and R⁸ are independently hydrogen, halo, hydroxy, cyano,alkyl or alkoxy;

R^(c) is hydrogen, cyano, halo, alkyl or alkoxy; and

n′ is 0, 1, 2, 3 or 4.

In some embodiments, R² is cyano, halo or alkyl. In some embodiments, R²is halo or alkyl. In some embodiments, R² is fluoro, chloro, bromo oriodo. In some embodiments, R² is fluoroalkyl. In some furtherembodiments, R² is —CH₂F, —CHF₂ or —CF₃.

In some embodiments, R³ is hydrogen, halo, cyano, alkyl, alkenyl,heteroalkyl or acyl; or R² and R³ taken together form a cyclic moiety.In a further embodiment, R³ is halo, cyano or alkyl. In yet a furtherembodiment, R³ is —(CH₂)_(n)OH, wherein n is 1, 2 or 3.

In some embodiments, R² and R³ taken together with the atoms to whichthey are attached form a 5- or 6-membered carbocycle with at least onesp³ hybridized carbon. Representative compounds with the carbocycleinclude, but are not limited to, the following:

wherein the carbocycle formed by linking R² and R³ may be optionallysubstituted with fluoro, chloro, hydroxy, alkyl or heteroalkyl. In afurther embodiment, the substituent(s) is selected from the groupconsisting of halo, C₁-C₄ alkyl, C₁-C₄ alkoxy and cyano. In yet otherembodiments, the substituent(s) is cycloalkyl or heterocycloalkyl andshares one or more ring atoms with the carbocycle formed by linking R²and R³. In some embodiments, the substituent(s) is C₃-C₅ cycloalkyl orC₃-C₅ heterocycloalkyl. In other embodiments, the substituent is oxo.

In some embodiments, R² and R³ taken together with the atoms to whichthey are attached form a 5- or 6-membered heterocycle, including, butnot limited to, a lactone or lactol, wherein said heterocycle may beoptionally substituted with fluoro, chloro, hydroxy, alkyl orheteroalkyl. In a further embodiment, the substituent(s) is selectedfrom the group consisting of halo, C₁-C₄ alkyl, C₁-C₄ alkoxy and cyano.

In some embodiments, R³ is hydrogen, R⁴ is —S(═O)₂R^(a) or—S(═O)(═NR^(b))R^(d), wherein R^(a) is fluoroalkyl, R^(b) is hydrogen,cyano or alkyl and R^(d) is alkyl.

In some embodiments, R⁴ is halo, cyano, fluoroalkyl, sulfinyl,sulfonamidyl, sulfonyl, sulfoximinyl or fluoroalkylsulfonyl. In someembodiments, R⁴ is cyano, fluoroalkyl, sulfonamidyl, sulfinyl, sulfonylor sulfoximinyl. In some embodiments, R⁴ is fluoroalkyl, sulfonamidyl,sulfonyl or sulfoximinyl. In a further embodiment, R⁴ is fluoroalkyl. Inyet another embodiment, R⁴ is sulfonyl. In still another embodiment, R⁴is alkylsulfonyl.

In some embodiments, R⁴ is —S(═O)₂R^(a), wherein R^(a) is alkyl orcycloalkyl. In a further embodiment, R^(a) is C₁-C₄ alkyl, optionallysubstituted with one or more fluorines. Suitable examples offluorine-substituted C₁-C₄ alkyl include, but are not limited to, —CH₂F,—CHF₂, —CF₃, —CH₂CF₃, —CH₂CHF₂, —CH₂CH₂F, —CHFCH₃ and —CF₂CH₃. In stilla further embodiment, R^(a) is methyl, optionally substituted with oneor more fluorines.

In some embodiments, R⁴ is —S(═O)(═NR^(b))R^(a), wherein R^(a) is alkylor cycloalkyl and R^(b) is hydrogen, cyano or alkyl. In a furtherembodiment, R^(a) is C₁-C₄ alkyl, optionally substituted with one ormore fluorines. Suitable examples of fluorine-substituted C₁-C₄ alkylinclude, but are not limited to, —CH₂F, —CHF₂, —CF₃, —CH₂CF₃, —CH₂CHF₂,—CH₂CH₂F, —CHFCH₃ and —CF₂CH₃.

In some embodiments, R⁴ is —S(═O)₂—N(R^(a))₂, wherein each R^(a) isindependently hydrogen, alkyl, heteroalkyl, cycloalkyl orheterocycloalkyl, and at least one R^(a) is hydrogen. In a furtherembodiment, both R^(a)s are hydrogen. In another further embodiment, oneR^(a) is hydrogen and the other R^(a) is C₁-C₄ alkyl.

In some embodiments, R⁴ is selected from the group consisting of —CN,—CF₃, —S(═O)CH₃, —S(═O)₂CH₃, —S(═O)₂CH₂F, —S(═O)₂CHF₂, —S(═O)₂CF₃,—S(═O)₂NH₂, —S(═O)₂NHCH₃, —S(═O)(═NH)CH₃, —S(═O)(═NH)CH₂F,—S(═O)(═NH)CHF₂, —S(═O)(═NH)CF₃, —S(═O)(═N—CN)CH₃, —S(═O)(═N—CN)CH₂F,—S(═O)(═N—CN)CHF₂ and —S(═O)(═N—CN)CF₃.

In some embodiments, R⁵ is hydrogen. In some other embodiments, R⁵ isC₁-C₄ alkyl or C₁-C₄ alkoxy. In a further embodiment, R⁵ is methyl.

In some embodiments, R⁶ is hydrogen. In some other embodiments, R⁶ isC₁-C₄ alkyl or C₁-C₄ alkoxy. In a further embodiment, R⁶ is methyl.

In some embodiments, R⁷ is hydrogen. In some other embodiments, R⁷ isC₁-C₄ alkyl or C₁-C₄ alkoxy. In a further embodiment, R⁷ is methyl.

In some embodiments, R⁸ is hydrogen. In some other embodiments, R⁸ isC₁-C₄ alkyl or C₁-C₄ alkoxy. In a further embodiment, R⁸ is methyl.

In some embodiments, R² and R³ taken together with the atoms to whichthey are attached form a 5- or 6-membered carbocycle with at least onesp³ hybridized carbon and R⁴ is cyano, fluoroalkyl, sulfonamidyl,sulfinyl, sulfonyl, sulfoximinyl or fluoroalkylsulfonyl. In a furtherembodiment, R⁴ is selected from the group consisting of —CN, —CF₃,—S(═O)CH₃, —S(═O)₂CH₃, —S(═O)₂CH₂F, —S(═O)₂CHF₂, —S(═O)₂CF₃, —S(═O)₂NH₂,—S(═O)₂NHCH₃, —S(═O)(═NH)CH₃, —S(═O)(═NH)CH₂F, —S(═O)(═NH)CHF₂,—S(═O)(═NH)CF₃, —S(═O)(═N—CN)CH₃, —S(═O)(═N—CN)CH₂F, —S(═O)(═N—CN)CHF₂and —S(═O)(═N—CN)CF₃.

In some embodiments, R³ is —CH₂OH and R⁴ is fluoroalkyl, sulfonamidyl,sulfonyl, sulfinyl or sulfoximinyl. In a further embodiment, R⁵ ishydrogen. In still a further embodiment, R² is cyano, halo or alkyl.

In some embodiments, R² is halo, cyano or alkyl; R³ is CH₂OH; R⁴ isfluoroalkyl, sulfonamidyl, sulfonyl, sulfinyl or sulfoximinyl. In afurther embodiment, R⁴ is selected from the group consisting of —CF₃,—S(═O)CH₃, —S(═O)₂CH₃, —S(═O)₂CH₂F, —S(═O)₂CHF₂, —S(═O)₂CF₃, —S(═O)₂NH₂,—S(═O)₂NHCH₃, —S(═O)(═NH)CH₃, —S(═O)(═NH)CH₂F, —S(═O)(═NH)CHF₂,—S(═O)(═NH)CF₃, —S(═O)(═N—CN)CH₃, —S(═O)(═N—CN)CH₂F, —S(═O)(═N—CN)CHF₂and —S(═O)(═N—CN)CF₃.

In some embodiments, X is N and Y is CR⁶. In other embodiments, X is CR⁵and Y is N. In still other embodiments, X is N and Y is N. In yet otherembodiments, X is CR⁵ and Y is CR⁶.

In some embodiments, Z is —O—, —S—, —S(O)—, —S(O)₂—, —S(O₂)N(R⁸)—,—C(O)—, —C(O)O—, —C(HR⁷)—, —N(R⁸)—, —C(O)N(R⁸)—, alkylene, alkenylene,alkynylene, heteroalkylene, or absent. In some embodiments, Z is —O—,—S—, —S(O)—, —S(O)₂—, —C(O)—, —C(HR⁷)—, —N(R⁸)—, C₁-C₃ alkylene, C₁-C₃heteroalkylene, C₁-C₃ alkenylene or absent. In some embodiments, Z is—O—. In other embodiments, Z is —S—. In further embodiments, Z is—C(HR⁷)—. In yet other embodiments, Z is —N(R⁸). In some embodiments, Zis absent.

In some embodiments, R^(c) is cyano, halo, C₁-C₄ alkyl or C₁-C₄ alkoxy.

In yet another aspect, the invention provides a compound of Formula I-C:

or a pharmaceutically acceptable salt or prodrug thereof, wherein:

X is CR⁵ or N;

Y is CR⁶ or N;

Z is —O—, —S—, —C(HR⁷)—, —N(R⁸)— or absent;

R¹ is alkyl, heteroalkyl, cycloalkyl, heterocycloalkyl, aryl,heteroaryl, acyl or cyano;

R⁴ is nitro, halo, cyano, alkyl, cycloalkyl, heteroaryl, carboxyl,sulfinyl, sulfonamidyl, sulfonyl or sulfoximinyl;

R⁵, R⁶, R⁷ and R⁸ are independently hydrogen, halo, hydroxy, cyano,alkyl or alkoxy;

R¹¹ is hydrogen, hydroxy, alkoxy or amino;

R¹² is hydrogen, alkyl, alkenyl or alkynyl; or R¹¹ and R¹² incombination form oxo or oxime;

each of R¹³ is independently selected from the group consisting ofhydrogen, fluoro, chloro, hydroxy, alkyl and heteroalkyl, with theproviso that when R¹³ is hydroxy, n is 1 or 2; or two R¹³s and thecarbon atom(s) to which they are attached form a 3- to 8-memberedcycloalkyl or heterocycloalkyl moiety; and

n is 0, 1, 2, 3 or 4.

In yet another aspect, the invention provides a compound of Formula I-C:

or a pharmaceutically acceptable salt or prodrug thereof, wherein:

X is CR⁵ or N;

Y is CR⁶ or N;

Z is —O—, —S—, —S(O)—, —S(O)₂—, —C(O)—, —C(HR⁷)—, —N(R⁸)—, C₁-C₃alkylene, C₁-C₃ heteroalkylene, C₁-C₃ alkenylene or absent;

R¹ is alkyl, heteroalkyl, cycloalkyl, heterocycloalkyl, aryl,heteroaryl, acyl or cyano;

R⁴ is nitro, halo, cyano, alkyl, cycloalkyl, heteroaryl, carboxyl,sulfinyl, sulfonamidyl, sulfonyl or sulfoximinyl;

R⁵, R⁶, R⁷ and R⁸ are independently hydrogen, halo, hydroxy, cyano,alkyl or alkoxy;

R¹¹ is hydrogen, halo, hydroxy, alkoxy or amino;

R¹² is hydrogen, alkyl, alkenyl or alkynyl; or R¹¹ and R¹² incombination form oxo or oxime;

each of R¹³ is independently selected from the group consisting ofhydrogen, fluoro, chloro, hydroxy, alkyl and heteroalkyl, with theproviso that when R¹³ is hydroxy, n is 1 or 2; or two R¹³s and thecarbon atom(s) to which they are attached form a 3- to 8-memberedcycloalkyl or heterocycloalkyl moiety; and

n is 0, 1, 2, 3 or 4.

In some embodiments, for a compound of Formula I-C, R¹ is furtherselected from alkyl, heteroalkyl, cycloalkyl, cycloalkenyl,heterocycloalkyl, aryl, heteroaryl, acyl and cyano.

In some embodiments, R¹ is phenyl or monocyclic heteroaryl. In somefurther embodiments, R¹ is phenyl or pyridyl, optionally substitutedwith one or more substituents selected from the group consisting ofhalo, alkyl, alkoxy and cyano. In a further embodiment, R¹ is

wherein the aryl ring is optionally substituted with one or moresubstituents selected from the group consisting of cyano, halo, alkyland alkoxy. In another further embodiment, R¹ is

wherein W¹ is N or CR¹⁰, R⁹ is cyano, halo, alkyl or alkoxy, and R¹⁰ ishydrogen, cyano, halo, alkyl or alkoxy.

In some embodiments, R¹ is bicyclic heteroaryl.

In some embodiments, R¹ is selected from the group consisting of:

and the rings specified for R¹ may optionally be substituted with one ormore substituents described for aryl and heteroaryl. In a furtherembodiment, the substituent(s) is selected from the group consisting ofhalo, C₁-C₄ alkyl, C₁-C₄ alkoxy and cyano.

In some embodiments, R¹ is selected from the group consisting of:

and the rings specified for R¹ may optionally be substituted with one ormore substituents described for cycloalkyl. In a further embodiment, thesubstituent(s) is selected from the group consisting of halo, hydroxy,C₁-C₄ alkyl, C₁-C₄ alkoxy, cyano and oxo.

In some embodiments, R¹ is cycloalkyl. In other embodiments, R¹ isheterocycloalkyl. In a further embodiment, R¹ is C₃-C₆ cycloalkyl orC₃-C₆ heterocycloalkyl. In yet a further embodiment, R¹ is cyclobutyl.In some embodiments, said cycloalkyl, cyclobutyl or heterocycloalkyl mayoptionally be substituted with one or more substituents described forcycloalkyl or heterocycloalkyl. In a further embodiment, thesubstituent(s) is selected from the group consisting of halo, C₁-C₄alkyl, C₁-C₄ alkoxy and cyano. In another further embodiment, thesubstituent(s) is at least one fluoro.

In some embodiments, R¹ is acyl or cyano. In a further embodiment, R¹ isacetyl.

In some embodiments, R¹ is alkyl. In a further embodiment, the alkyl issubstituted with at least one substituent(s) selected from the groupconsisting of halo, C₁-C₄ alkyl, C₁-C₄ alkoxy and cyano. In anotherfurther embodiment, the alkyl is substituted with at least one fluoro.In some embodiments, R¹ is heteroalkyl.

In some embodiments, R¹ is selected from the group consisting of:

wherein each of the members may optionally be substituted with one ormore substituents selected from the group consisting of cyano, halo,alkyl and alkoxy. In a further embodiment, the substituent(s) isselected from the group consisting of fluoro, C₁-C₄ alkyl, C₁-C₄ alkoxyand cyano.

In some embodiments, R⁴ is cyano, fluoroalkyl, sulfinyl, sulfonamidyl,sulfonyl or sulfoximinyl. In some embodiments, R⁴ is fluoroalkyl,sulfonamidyl, sulfinyl, sulfonyl or sulfoximinyl. In a furtherembodiment, R⁴ is fluoroalkyl. In yet another embodiment, R⁴ issulfonyl. In still another embodiment, R⁴ is alkylsulfonyl.

In some embodiments, R⁴ is —S(═O)₂R^(a), wherein R^(a) is alkyl orcycloalkyl. In a further embodiment, R^(a) is C₁-C₄ alkyl, optionallysubstituted with one or more fluorines. Suitable examples offluorine-substituted C₁-C₄ alkyl include, but are not limited to, —CH₂F,—CHF₂, —CF₃, —CH₂CF₃, —CH₂CHF₂, —CH₂CH₂F, —CHFCH₃ and —CF₂CH₃. In stilla further embodiment, R^(a) is methyl, optionally substituted with oneor more fluorines.

In some embodiments, R⁴ is —S(═O)(═NR^(b))R^(a), wherein R^(a) is alkylor cycloalkyl and R^(b) is hydrogen, cyano or alkyl. In a furtherembodiment, R^(a) is C₁-C₄ alkyl, optionally substituted with one ormore fluorines. Suitable examples of fluorine-substituted C₁-C₄ alkylinclude, but are not limited to, —CH₂F, —CHF₂, —CF₃, —CH₂CF₃, —CH₂CHF₂,—CH₂CH₂F, —CHFCH₃ and —CF₂CH₃.

In some embodiments, R⁴ is —S(═O)₂N(R^(a))₂, wherein each R^(a) isindependently hydrogen, alkyl, heteroalkyl, cycloalkyl orheterocycloalkyl, and at least one R^(a) is hydrogen. In another furtherembodiment, one R^(a) is hydrogen and the other R^(a) is C₁-C₄ alkyl.

In some embodiments, R⁴ is selected from the group consisting of —CN,—CF₃, —S(═O)CH₃, —S(═O)₂CH₃, —S(═O)₂CH₂F, —S(═O)₂CHF₂, —S(═O)₂CF₃,—S(═O)₂NH₂, —S(═O)₂NHCH₃, —S(═O)(═NH)CH₃, —S(═O)(═NH)CH₂F,—S(═O)(═NH)CHF₂, —S(═O)(═NH)CF₃, —S(═O)(═N—CN)CH₃, —S(═O)(═N—CN)CH₂F,—S(═O)(═N—CN)CHF₂ and —S(═O)(═N—CN)CF₃.

In some embodiments, R⁵ is hydrogen or alkyl. In some other embodiments,R⁵ is C₁-C₄ alkyl or C₁-C₄ alkoxy. In a further embodiment, R⁵ ismethyl.

In some embodiments, R⁶ is hydrogen or alkyl. In some other embodiments,R⁶ is C₁-C₄ alkyl or C₁-C₄ alkoxy. In a further embodiment, R⁶ ismethyl.

In some embodiments, R⁷ is hydrogen or alkyl. In some other embodiments,R⁷ is C₁-C₄ alkyl or C₁-C₄ alkoxy. In a further embodiment, R⁷ ismethyl.

In some embodiments, R⁸ is hydrogen or alkyl. In some other embodiments,R⁸ is C₁-C₄ alkyl or C₁-C₄ alkoxy. In a further embodiment, R⁸ ismethyl.

In some embodiments, R¹¹ is hydroxy or amino. In a further embodiment,R¹¹ is hydroxy. In another further embodiment, R¹¹ is amino.

In some embodiments, R¹² is hydrogen. In some other embodiments, R¹² isalkyl or alkenyl.

In some embodiments, R¹³ is fluoro. In a further embodiment, n is 1, 2or 3. In a further embodiment, two R¹³s in combination form oxo, oximeor methylene. In still further embodiments, two R¹³s and the carbonatom(s) to which they are attached form a 3- to 8-membered cycloalkyl orheterocycloalkyl moiety.

In some embodiments, R¹ is monocyclic aryl or monocyclic heteroaryl andR¹¹ is hydroxy or amino. In a further embodiment, R¹³ is fluoro. Instill a further embodiment, n is 1, 2 or 3.

In some embodiments, R¹ is phenyl or monocyclic heteroaryl, R¹¹ ishydroxy or amino, R¹³ is fluoro, n is 1, 2 or 3, and R⁵ is hydrogen.

In some embodiments, R¹ is bicyclic heteroaryl and R¹¹ is hydroxy oramino. In a further embodiment, R¹³ is fluoro. In still a furtherembodiment, n is 1, 2 or 3.

In some embodiments, R¹ is bicyclic heteroaryl, R¹¹ is hydroxy or amino,R¹³ is fluoro, n is 1, 2 or 3, and R⁵ is hydrogen.

In some embodiments, R⁴ is cyano, fluoroalkyl, sulfonamidyl, sulfinyl,sulfonyl, sulfoximinyl or fluoroalkylsulfonyl, and R¹¹ is hydroxy oramino. In a further embodiment, R¹² is hydrogen. In another furtherembodiment, R¹³ is fluoro. In still a further embodiment, n is 1, 2 or3.

In some embodiments, R⁴ is cyano, fluoroalkyl, sulfonamidyl, sulfinyl,sulfonyl, sulfoximinyl or fluoroalkylsulfonyl; R¹¹ is hydroxy or amino;R¹³ is fluoro; n is 1, 2 or 3; and R⁵ is hydrogen. In a furtherembodiment, R¹² is hydrogen.

In some embodiments R¹¹ is hydroxy or amino and R¹² is hydrogen. In afurther embodiment, R¹³ is fluoro. In still a further embodiment, n is1, 2 or 3.

In some embodiments R¹¹ is hydroxy or amino, R¹² is hydrogen, R¹³ isfluoro, n is 1, 2 or 3, and R⁵ is hydrogen. In a further embodiment, R⁴is selected from the group consisting of —CN, —CF₃, —S(═O)CH₃,—S(═O)₂CH₃, —S(═O)₂CH₂F, —S(═O)₂CHF₂, —S(═O)₂CF₃, —S(═O)₂NH₂,—S(═O)₂NHCH₃, —S(═O)(═NH)CH₃, —S(═O)(═NH)CH₂F, —S(═O)(═NH)CHF₂,—S(═O)(═NH)CF₃, —S(═O)(═N—CN)CH₃, —S(═O)(═N—CN)CH₂F, —S(═O)(═N—CN)CHF₂and —S(═O)(═N—CN)CF₃.

In some embodiments, R⁴ is fluoroalkyl; n is 0, 1, 2 or 3; Z is —O—; R¹¹is hydroxy; and R¹² is hydrogen.

In some embodiments, R⁴ is sulfonyl or fluoroalkylsulfonyl; n is 0, 1, 2or 3; Z is —O—; R¹¹ is hydroxy; and R¹² is hydrogen.

In some embodiments, X is N and Y is CR⁶. In other embodiments, X is CR⁵and Y is N. In still other embodiments, X is N and Y is N. In yet otherembodiments, X is CR⁵ and Y is CR⁶.

In some embodiments, Z is —O—, —S—, —S(O)—, —S(O)₂—, —S(O₂)N(R⁸)—,—C(O)—, —C(O)O—, —C(HR⁷)—, —N(R⁸)—, —C(O)N(R⁸)—, alkylene, alkenylene,alkynylene, heteroalkylene, or absent. In some embodiments, Z is —O—,—S—, —S(O)—, —S(O)₂—, —C(O)—, —C(HR⁷)—, —N(R⁸)—, C₁-C₃ alkylene, C₁-C₃heteroalkylene, C₁-C₃ alkenylene or absent. In some embodiments, Z is—O—. In other embodiments, Z is —S—. In further embodiments, Z is—C(HR⁷)—. In yet other embodiments, Z is —N(R⁸). In some embodiments, Zis absent.

In still another aspect, the invention provides a compound of FormulaI-D, I-E, I-F or I-G:

or a pharmaceutically acceptable salt or prodrug thereof, wherein:

X is CR⁵ or N;

Y is CR⁶ or N;

Z is —O—, —S—, —C(HR⁷)—, —N(R⁸)—, or absent;

R¹ is alkyl, heteroalkyl, cycloalkyl, heterocycloalkyl, aryl orheteroaryl;

R⁴ is nitro, halo, cyano, alkyl, cycloalkyl, heteroaryl, carboxyl,sulfinyl, sulfonamidyl, sulfonyl or sulfoximinyl;

R⁵, R⁶, R⁷ and R⁸ are independently hydrogen, halo, hydroxy, cyano,alkyl or alkoxy; and

R¹¹ is hydrogen, hydroxy, alkoxy or amino.

In still another aspect, the invention provides a compound of FormulaI-D, I-E, I-F or I-G:

or a pharmaceutically acceptable salt or prodrug thereof, wherein:

X is CR⁵ or N;

Y is CR⁶ or N;

Z is —O—, —S—, —S(O)—, —S(O)₂—, —C(O)—, —C(HR⁷)—, —N(R⁸)—, C₁-C₃alkylene, C₁-C₃ heteroalkylene, C₁-C₃ alkenylene or absent;

R¹ is alkyl, heteroalkyl, cycloalkyl, heterocycloalkyl, aryl orheteroaryl;

R⁴ is nitro, halo, cyano, alkyl, cycloalkyl, heteroaryl, carboxyl,sulfinyl, sulfonamidyl, sulfonyl or sulfoximinyl;

R⁵, R⁶, R⁷ and R⁸ are independently hydrogen, halo, hydroxy, cyano,alkyl or alkoxy; and

R¹¹ is hydrogen, halo, hydroxy, alkoxy or amino.

In some embodiments, for a compound of Formula I-D, I-E, I-F, or I-G, R¹is further selected from alkyl, heteroalkyl, cycloalkyl, cycloalkenyl,heterocycloalkyl, aryl and heteroaryl.

In some embodiments, R¹ is monocyclic aryl or monocyclic heteroaryl. Insome further embodiments, R is phenyl or pyridyl, optionally substitutedwith one or more substituents selected from the group consisting ofhalo, alkyl, alkoxy and cyano. In a further embodiment, R¹ is

wherein the aryl ring is optionally substituted with one or moresubstituents selected from the group consisting of cyano, halo, alkyland alkoxy. In another further embodiment, R¹ is

wherein W¹ is N or CR¹⁰, R⁹ is cyano, halo, alkyl or alkoxy, and R¹⁰ ishydrogen, cyano, halo, alkyl or alkoxy.

In some embodiments, R¹ is bicyclic heteroaryl having at least one Natom.

In some embodiments, R¹ is selected from the group consisting of:

and the rings specified for R¹ may optionally be substituted with one ormore substituents described for aryl and heteroaryl. In a furtherembodiment, the substituent(s) is selected from the group consisting ofhalo, C₁-C₄ alkyl, C₁-C₄ alkoxy and cyano.

In some embodiments, R¹ is selected from the group consisting of:

and the rings specified for R¹ may optionally be substituted with one ormore substituents described for cycloalkyl. In a further embodiment, thesubstituent(s) is selected from the group consisting of halo, hydroxy,C₁-C₄ alkyl, C₁-C₄ alkoxy, cyano and oxo.

In some embodiments, R¹ is cycloalkyl. In other embodiments, R¹ isheterocycloalkyl. In a further embodiment, R¹ is C₃-C₆ cycloalkyl orC₃-C₆ heterocycloalkyl. In yet a further embodiment, R¹ is cyclobutyl.In some embodiments, said cycloalkyl, cyclobutyl or heterocycloalkyl mayoptionally be substituted with one or more substituents described forcycloalkyl or heterocycloalkyl. In a further embodiment, thesubstituent(s) is selected from the group consisting of halo, C₁-C₄alkyl, C₁-C₄ alkoxy and cyano. In another further embodiment, thesubstituent(s) is at least one fluoro.

In some embodiments, R¹ is acyl or cyano. In a further embodiment, R¹ isacetyl.

In some embodiments, R¹ is alkyl. In a further embodiment, the alkyl issubstituted with at least one substituent(s) selected from the groupconsisting of halo, C₁-C₄ alkyl, C₁-C₄ alkoxy and cyano. In anotherfurther embodiment, the alkyl is substituted with at least one fluoro.In some embodiments, R¹ is heteroalkyl.

In some embodiments, R¹ is selected from the group consisting of:

wherein each of the members may optionally be substituted with one ormore substituents selected from the group consisting of cyano, halo,alkyl and alkoxy. In a further embodiment, the substituent(s) isselected from the group consisting of fluoro, C₁-C₄ alkyl, C₁-C₄ alkoxyand cyano.

In some embodiments, R⁴ is cyano, fluoroalkyl, sulfinyl, sulfonamidyl,sulfonyl or sulfoximinyl. In some embodiments, R⁴ is fluoroalkyl,sulfonamidyl, sulfinyl, sulfonyl or sulfoximinyl. In a furtherembodiment, R⁴ is fluoroalkyl. In yet another embodiment, R⁴ issulfonyl. In still another embodiment, R⁴ is alkylsulfonyl.

In some embodiments, R⁴ is —S(═O)₂R^(a), wherein R^(a) is alkyl orcycloalkyl. In a further embodiment, R^(a) is C₁-C₄ alkyl, optionallysubstituted with one or more fluorines. Suitable examples offluorine-substituted C₁-C₄ alkyl include, but are not limited to, —CH₂F,—CHF₂, —CF₃, —CH₂CF₃, —CH₂CHF₂, —CH₂CH₂F, —CHFCH₃ and —CF₂CH₃.

In some embodiments, R⁴ is —S(═O)(═NR^(b))R^(a), wherein R^(a) is alkylor cycloalkyl and R^(b) is hydrogen, cyano or alkyl. In a furtherembodiment, R^(a) is C₁-C₄ alkyl, optionally substituted with one ormore fluorines. Suitable examples of fluorine-substituted C₁-C₄ alkylinclude, but are not limited to, —CH₂F, —CHF₂, —CF₃, —CH₂CF₃, —CH₂CHF₂,—CH₂CH₂F, —CHFCH₃ and —CF₂CH₃.

In some embodiments, R⁴ is —S(═O)₂N(R^(a))₂, wherein each R^(a) isindependently hydrogen, alkyl, heteroalkyl, cycloalkyl orheterocycloalkyl, and at least one R^(a) is hydrogen. In another furtherembodiment, both R^(a)s are hydrogen. In a further embodiment, one R^(a)is hydrogen and the other R^(a) is C₁-C₄ alkyl.

In some embodiments, R⁴ is selected from the group consisting of —CN,—CF₃, —S(═O)CH₃, —S(═O)₂CH₃, —S(═O)₂CH₂F, —S(═O)₂CHF₂, —S(═O)₂CF₃,—S(═O)₂NH₂, —S(═O)₂NHCH₃, —S(═O)(═NH)CH₃, —S(═O)(═NH)CH₂F,—S(═O)(═NH)CHF₂, —S(═O)(═NH)CF₃, —S(═O)(═N—CN)CH₃, —S(═O)(═N—CN)CH₂F,—S(═O)(═N—CN)CHF₂ and —S(═O)(═N—CN)CF₃.

In some embodiments, R⁵ is hydrogen or alkyl. In some other embodiments,R⁵ is C₁-C₄ alkyl or C₁-C₄ alkoxy. In a further embodiment, R⁵ ismethyl.

In some embodiments, R⁶ is hydrogen or alkyl. In some other embodiments,R⁶ is C₁-C₄ alkyl or C₁-C₄ alkoxy. In a further embodiment, R⁶ ismethyl.

In some embodiments, R⁷ is hydrogen or alkyl. In some other embodiments,R⁷ is C₁-C₄ alkyl or C₁-C₄ alkoxy. In a further embodiment, R⁷ ismethyl.

In some embodiments, R⁸ is hydrogen or alkyl. In some other embodiments,R⁸ is C₁-C₄ alkyl or C₁-C₄ alkoxy. In a further embodiment, R⁸ ismethyl.

In some embodiments, R¹¹ is hydroxy. In another further embodiment, R¹¹is amino.

In some embodiments, R¹ is phenyl or monocyclic heteroaryl and R¹¹ ishydroxy or amino.

In some embodiments, R¹ is bicyclic heteroaryl and R¹¹ is hydroxy oramino. In a further embodiment, X is CR⁵ and R⁵ is hydrogen. In anotherfurther embodiment, R⁵ is alkyl. In still a further embodiment, R⁵ isC₁-C₄ alkyl.

In some embodiments R¹ is bicyclic heteroaryl and R⁴ is cyano,fluoroalkyl, sulfinyl, sulfonamidyl, sulfonyl or sulfoximinyl. In afurther embodiment, X is CR⁵ and R⁵ is hydrogen. In another furtherembodiment, R⁴ is selected from the group consisting of —CN, —CF₃,—S(═O)CH₃, —S(═O)₂CH₃, —S(═O)₂CH₂F, —S(═O)₂CHF₂, —S(═O)₂CF₃, —S(═O)₂NH₂,—S(═O)₂NHCH₃, —S(═O)(═NH)CH₃, —S(═O)(═NH)CH₂F, —S(═O)(═NH)CHF₂,—S(═O)(═NH)CF₃, —S(═O)(═N—CN)CH₃, —S(═O)(═N—CN)CH₂F, —S(═O)(═N—CN)CHF₂and —S(═O)(═N—CN)CF₃.

In some embodiments R¹ is bicyclic heteroaryl; R⁴ is cyano, fluoroalkyl,sulfinyl, sulfonamidyl, sulfonyl or sulfoximinyl; R¹¹ is hydroxy oramino; and X is CR⁵ or N; and R⁵ is hydrogen. In a further embodiment,R¹¹ is hydroxy. In another further embodiment, R⁴ is selected from thegroup consisting of —CN, —CF₃, —S(═O)CH₃, —S(═O)₂CH₃, —S(═O)₂CH₂F,—S(═O)₂CHF₂, —S(═O)₂CF₃, —S(═O)₂NH₂, —S(═O)₂NHCH₃, —S(═O)(═NH)CH₃,—S(═O)(═NH)CH₂F, —S(═O)(═NH)CHF₂, —S(═O)(═NH)CF₃, —S(═O)(═N—CN)CH₃,—S(═O)(═N—CN)CH₂F, —S(═O)(═N—CN)CHF₂ and —S(═O)(═N—CN)CF₃.

In some embodiments R¹ is phenyl or monocyclic heteroaryl and R⁴ iscyano, fluoroalkyl, sulfinyl, sulfonamidyl, sulfonyl or sulfoximinyl. Ina further embodiment, X is CR⁵ and R⁵ is hydrogen. In another furtherembodiment, R⁴ is selected from the group consisting of —CN, —CF₃,—S(═O)CH₃, —S(═O)₂CH₃, —S(═O)₂CH₂F, —S(═O)₂CHF₂, —S(═O)₂CF₃, —S(═O)₂NH₂,—S(═O)₂NHCH₃, —S(═O)(═NH)CH₃, —S(═O)(═NH)CH₂F, —S(═O)(═NH)CHF₂,—S(═O)(═NH)CF₃, —S(═O)(═N—CN)CH₃, —S(═O)(═N—CN)CH₂F, —S(═O)(═N—CN)CHF₂and —S(═O)(═N—CN)CF₃.

In some embodiments R¹ is phenyl or monocyclic heteroaryl; R⁴ is cyano,fluoroalkyl, sulfinyl, sulfonamidyl, sulfonyl or sulfoximinyl; R¹¹ ishydroxy or amino; and X is CR⁵ or N; and R⁵ is hydrogen. In a furtherembodiment, R¹¹ is hydroxy. In another further embodiment, R⁴ isselected from the group consisting of —CN, —CF₃, —S(═O)CH₃, —S(═O)₂CH₃,—S(═O)₂CH₂F, —S(═O)₂CHF₂, —S(═O)₂CF₃, —S(═O)₂NH₂, —S(═O)₂NHCH₃,—S(═O)(═NH)CH₃, —S(═O)(═NH)CH₂F, —S(═O)(═NH)CHF₂, —S(═O)(═NH)CF₃,—S(═O)(═N—CN)CH₃, —S(═O)(═N—CN)CH₂F, —S(═O)(═N—CN)CHF₂ and—S(═O)(═N—CN)CF₃.

In some embodiments, X is N and Y is CR⁶. In other embodiments, X is CR⁵and Y is N. In still other embodiments, X is N and Y is N. In yet otherembodiments, X is CR⁵ and Y is CR⁶.

In some embodiments, Z is —O—, —S—, —S(O)—, —S(O)₂—, —S(O₂)N(R⁸)—,—C(O)—, —C(O)O—, —C(HR⁷)—, —N(R⁸)—, —C(O)N(R⁸)—, alkylene, alkenylene,alkynylene, heteroalkylene, or absent. In some embodiments, Z is —O—,—S—, —S(O)—, —S(O)₂—, —C(O)—, —C(HR⁷)—, —N(R⁸)—, C₁-C₃ alkylene, C₁-C₃heteroalkylene, C₁-C₃ alkenylene or absent. In some embodiments, Z is—O—. In other embodiments, Z is —S—. In further embodiments, Z is—C(HR⁷)—. In yet other embodiments, Z is —N(R⁸). In some embodiments, Zis absent.

In a further aspect, the invention provides a compound of Formula I-H,I-I, I-J or I-K:

or a pharmaceutically acceptable salt or prodrug thereof, wherein:

X is CR⁵ or N;

Y is CR⁶ or N;

Z is —O—, —S—, —C(HR⁷)—, —N(R⁸)— or absent;

R¹ is alkyl, heteroalkyl, cycloalkyl, heterocycloalkyl, aryl orheteroaryl;

R⁴ is nitro, halo, cyano, alkyl, cycloalkyl, heteroaryl, carboxyl,sulfinyl, sulfonamidyl, sulfonyl or sulfoximinyl;

R⁵, R⁶, R⁷ and R⁸ are independently hydrogen, halo, hydroxy, cyano,alkyl or alkoxy; and

R¹¹ is hydroxy or amino.

In a further aspect, the invention provides a compound of Formula I-H,I-I, I-J or I-K:

or a pharmaceutically acceptable salt or prodrug thereof, wherein:

X is CR⁵ or N;

Y is CR⁶ or N;

Z is —O—, —S—, —S(O)—, —S(O)₂—, —C(O)—, —C(HR⁷)—, —N(R⁸)—, C₁-C₃alkylene, C₁-C₃ heteroalkylene, C₁-C₃ alkenylene or absent;

R¹ is alkyl, heteroalkyl, cycloalkyl, heterocycloalkyl, aryl orheteroaryl;

R⁴ is nitro, halo, cyano, alkyl, cycloalkyl, heteroaryl, carboxyl,sulfinyl, sulfonamidyl, sulfonyl or sulfoximinyl;

R⁵, R⁶, R⁷ and R⁸ are independently hydrogen, halo, hydroxy, cyano,alkyl or alkoxy; and

R¹¹ is hydroxy or amino.

In some embodiments, for a compound of Formula I-H, I-I, I-J, or I-K, R¹is further selected from alkyl, heteroalkyl, cycloalkyl, cycloalkenyl,heterocycloalkyl, aryl and heteroaryl.

In some embodiments, R¹ is monocyclic aryl or monocyclic heteroaryl. Insome further embodiments, R is phenyl or pyridyl, optionally substitutedwith one or more substituents selected from the group consisting ofhalo, alkyl, alkoxy and cyano. In a further embodiment, R¹ is

wherein the aryl ring is optionally substituted with one or moresubstituents selected from the group consisting of cyano, halo, alkyland alkoxy. In another further embodiment, R¹ is

wherein W¹ is N or CR¹⁰, R⁹ is cyano, halo, alkyl or alkoxy, and R¹⁰ ishydrogen, cyano, halo, alkyl or alkoxy.

In some embodiments, R¹ is bicyclic heteroaryl.

In some embodiments, R¹ is selected from the group consisting of:

and the rings specified for R¹ may optionally be substituted with one ormore substituents described for aryl and heteroaryl. In a furtherembodiment, the substituent(s) is selected from the group consisting ofhalo, C₁-C₄ alkyl, C₁-C₄ alkoxy and cyano.

In some embodiments, R¹ is selected from the group consisting of:

and the rings specified for R¹ may optionally be substituted with one ormore substituents described for cycloalkyl. In a further embodiment, thesubstituent(s) is selected from the group consisting of halo, hydroxy,C₁-C₄ alkyl, C₁-C₄ alkoxy, cyano and oxo.

In some embodiments, R¹ is cycloalkyl. In other embodiments, R¹ isheterocycloalkyl. In a further embodiment, R¹ is C₃-C₆ cycloalkyl orC₃-C₆ heterocycloalkyl. In yet a further embodiment, R¹ is cyclobutyl.In some embodiments, said cycloalkyl, cyclobutyl or heterocycloalkyl mayoptionally be substituted with one or more substituents described forcycloalkyl or heterocycloalkyl. In a further embodiment, thesubstituent(s) is selected from the group consisting of halo, C₁-C₄alkyl, C₁-C₄ alkoxy and cyano. In another further embodiment, thesubstituent(s) is at least one fluoro.

In some embodiments, R¹ is acyl or cyano. In a further embodiment, R¹ isacetyl.

In some embodiments, R¹ is alkyl. In a further embodiment, the alkyl issubstituted with at least one substituent(s) selected from the groupconsisting of halo, C₁-C₄ alkyl, C₁-C₄ alkoxy and cyano. In anotherfurther embodiment, the alkyl is substituted with at least one fluoro.In some embodiments, R¹ is heteroalkyl.

In some embodiments, R¹ is alkyl. In a further embodiment, the alkyl issubstituted with at least one substituent(s) selected from the groupconsisting of halo, C₁-C₄ alkyl, C₁-C₄ alkoxy and cyano. In anotherfurther embodiment, the alkyl is substituted with at least one fluoro.In some embodiments, R¹ is heteroalkyl.

In some embodiments, R¹ is selected from the group consisting of:

wherein each of the members may optionally be substituted with one ormore substituents selected from the group consisting of cyano, halo,alkyl and alkoxy. In a further embodiment, the substituent(s) isselected from the group consisting of fluoro, C₁-C₄ alkyl, C₁-C₄ alkoxyand cyano.

In some embodiments, R⁴ is cyano, fluoroalkyl, sulfinyl, sulfonamidyl,sulfonyl or sulfoximinyl. In a further embodiment, R⁴ is fluoroalkyl. Inyet another embodiment, R⁴ is sulfonyl. In still another embodiment, R⁴is alkylsulfonyl.

In some embodiments, R⁴ is —S(═O)₂R^(a), wherein R^(a) is alkyl orcycloalkyl. In a further embodiment, R^(a) is C₁-C₄ alkyl, optionallysubstituted with one or more fluorines. Suitable examples offluorine-substituted C₁-C₄ alkyl include, but are not limited to, —CH₂F,—CHF₂, —CF₃, —CH₂CF₃, —CH₂CHF₂, —CH₂CH₂F, —CHFCH₃ and —CF₂CH₃. In stilla further embodiment, R^(a) is methyl, optionally substituted with oneor more fluorines.

In some embodiments, R⁴ is —S(═O)(═NR^(b))R^(a), wherein R^(a) is alkylor cycloalkyl and R^(b) is hydrogen, cyano or alkyl. In a furtherembodiment, R^(a) is C₁-C₄ alkyl, optionally substituted with one ormore fluorines. Suitable examples of fluorine-substituted C₁-C₄ alkylinclude, but are not limited to, —CH₂F, —CHF₂, —CF₃, —CH₂CF₃, —CH₂CHF₂,—CH₂CH₂F, —CHFCH₃ and —CF₂CH₃.

In some embodiments, R⁴ is —S(═O)₂N(R^(a))₂, wherein each R^(a) isindependently hydrogen, alkyl, heteroalkyl, cycloalkyl orheterocycloalkyl, and at least one R^(a) is hydrogen. In another furtherembodiment, both R^(a)s are hydrogen. In a further embodiment, one R^(a)is hydrogen and the other R^(a) is C₁-C₄ alkyl.

In some embodiments, R⁴ is selected from the group consisting of —CN,—CF₃, —S(═O)CH₃, —S(═O)₂CH₃, —S(═O)₂CH₂F, —S(═O)₂CHF₂, —S(═O)₂CF₃,—S(═O)₂NH₂, —S(═O)₂NHCH₃, —S(═O)(═NH)CH₃, —S(═O)(═NH)CH₂F,—S(═O)(═NH)CHF₂, —S(═O)(═NH)CF₃, —S(═O)(═N—CN)CH₃, —S(═O)(═N—CN)CH₂F,—S(═O)(═N—CN)CHF₂ and —S(═O)(═N—CN)CF₃.

In some embodiments, R⁵ is hydrogen or alkyl. In some other embodiments,R⁵ is C₁-C₄ alkyl or C₁-C₄ alkoxy. In a further embodiment, R⁵ ismethyl.

In some embodiments, R⁶ is hydrogen or alkyl. In some other embodiments,R⁶ is C₁-C₄ alkyl or C₁-C₄ alkoxy. In a further embodiment, R⁶ ismethyl.

In some embodiments, R⁷ is hydrogen or alkyl. In some other embodiments,R⁷ is C₁-C₄ alkyl or C₁-C₄ alkoxy. In a further embodiment, R⁷ ismethyl.

In some embodiments, R⁸ is hydrogen or alkyl. In some other embodiments,R⁸ is C₁-C₄ alkyl or C₁-C₄ alkoxy. In a further embodiment, R⁸ ismethyl.

In some embodiments, R¹¹ is hydroxy. In another further embodiment, R¹¹is amino.

In some embodiments, R¹ is phenyl or monocyclic heteroaryl and R¹¹ ishydroxy or amino.

In some embodiments, R¹ is bicyclic heteroaryl and R¹¹ is hydroxy oramino. In a further embodiment, X is CR⁵ and R⁵ is hydrogen. In anotherfurther embodiment, R⁵ is alkyl. In still a further embodiment, R⁵ isC₁-C₄ alkyl.

In some embodiments R¹ is bicyclic heteroaryl and R⁴ is cyano,fluoroalkyl, sulfinyl, sulfonamidyl, sulfonyl or sulfoximinyl. In afurther embodiment, X is CR⁵ and R⁵ is hydrogen. In another furtherembodiment, R⁴ is selected from the group consisting of —CN, —CF₃,—S(═O)CH₃, —S(═O)₂CH₃, —S(═O)₂CH₂F, —S(═O)₂CHF₂, —S(═O)₂CF₃, —S(═O)₂NH₂,—S(═O)₂NHCH₃, —S(═O)(═NH)CH₃, —S(═O)(═NH)CH₂F, —S(═O)(═NH)CHF₂,—S(═O)(═NH)CF₃, —S(═O)(═N—CN)CH₃, —S(═O)(═N—CN)CH₂F, —S(═O)(═N—CN)CHF₂and —S(═O)(═N—CN)CF₃.

In some embodiments R¹ is bicyclic heteroaryl; R⁴ is cyano, fluoroalkyl,sulfinyl, sulfonamidyl, sulfonyl or sulfoximinyl; R¹¹ is hydroxy oramino; and X is CR⁵ or N; and R⁵ is hydrogen. In a further embodiment,R¹¹ is hydroxy. In another further embodiment, R⁴ is selected from thegroup consisting of —CN, —CF₃, —S(═O)CH₃, —S(═O)₂CH₃, —S(═O)₂CH₂F,—S(═O)₂CHF₂, —S(═O)₂CF₃, —S(═O)₂NH₂, —S(═O)₂NHCH₃, —S(═O)(═NH)CH₃,—S(═O)(═NH)CH₂F, —S(═O)(═NH)CHF₂, —S(═O)(═NH)CF₃, —S(═O)(═N—CN)CH₃,—S(═O)(═N—CN)CH₂F, —S(═O)(═N—CN)CHF₂ and —S(═O)(═N—CN)CF₃.

In some embodiments R¹ is phenyl or monocyclic heteroaryl and R⁴ iscyano, fluoroalkyl, sulfinyl, sulfonamidyl, sulfonyl or sulfoximinyl. Ina further embodiment, X is CR⁵ and R⁵ is hydrogen. In another furtherembodiment, R⁴ is selected from the group consisting of —CN, —CF₃,—S(═O)CH₃, —S(═O)₂CH₃, —S(═O)₂CH₂F, —S(═O)₂CHF₂, —S(═O)₂CF₃, —S(═O)₂NH₂,—S(═O)₂NHCH₃, —S(═O)(═NH)CH₃, —S(═O)(═NH)CH₂F, —S(═O)(═NH)CHF₂,—S(═O)(═NH)CF₃, —S(═O)(═N—CN)CH₃, —S(═O)(═N—CN)CH₂F, —S(═O)(═N—CN)CHF₂and —S(═O)(═N—CN)CF₃.

In some embodiments R¹ is phenyl or monocyclic heteroaryl; R⁴ is cyano,fluoroalkyl, sulfinyl, sulfonamidyl, sulfonyl or sulfoximinyl; R¹¹ ishydroxy or amino; and X is CR⁵ or N; and R⁵ is hydrogen. In a furtherembodiment, R¹¹ is hydroxy. In another further embodiment, R⁴ isselected from the group consisting of —CN, —CF₃, —S(═O)CH₃, —S(═O)₂CH₃,—S(═O)₂CH₂F, —S(═O)₂CHF₂, —S(═O)₂CF₃, —S(═O)₂NH₂, —S(═O)₂NHCH₃,—S(═O)(═NH)CH₃, —S(═O)(═NH)CH₂F, —S(═O)(═NH)CHF₂, —S(═O)(═NH)CF₃,—S(═O)(═N—CN)CH₃, —S(═O)(═N—CN)CH₂F, —S(═O)(═N—CN)CHF₂ and—S(═O)(═N—CN)CF₃.

In some embodiments, X is N and Y is CR⁶. In other embodiments, X is CR⁵and Y is N. In still other embodiments, X is N and Y is N. In yet otherembodiments, X is CR⁵ and Y is CR⁶.

In some embodiments, Z is —O—, —S—, —S(O)—, —S(O)₂—, —S(O₂)N(R⁸)—,—C(O)—, —C(O)O—, —C(HR⁷)—, —N(R⁸)—, —C(O)N(R⁸)—, alkylene, alkenylene,alkynylene, heteroalkylene, or absent. In some embodiments, Z is —O—,—S—, —S(O)—, —S(O)₂—, —C(O)—, —C(HR⁷)—, —N(R⁸)—, C₁-C₃ alkylene, C₁-C₃heteroalkylene, C₁-C₃ alkenylene or absent. In some embodiments, Z is—O—. In other embodiments, Z is —S—. In further embodiments, Z is—C(HR⁷)—. In yet other embodiments, Z is —N(R⁸). In some embodiments, Zis absent.

In some embodiments, a compound of any one of Formulae I-H-I-K may havean enantiomeric excess of at least about 80%, at least about 81%, atleast about 82%, at least about 83%, at least about 84%, at least about85%, at least about 86%, at least about 87%, at least about 88%, atleast about 89%, at least about 90%, at least about 91%, at least about92%, at least about 93%, at least about 94%, at least about 95%, atleast about 96%, at least about 97%, at least about 98%, or even higher.In some embodiments, a compound of any one of Formulae I-H-I-K may havean enantiomeric excess of about 80%, about 81%, about 82%, about 83%,about 84%, about 85%, about 86%, about 87%, about 88%, about 89%, about90%, about 91%, about 92%, about 93%, about 94%, about 95%, about 96%,about 97%, about 98% or about 99%.

In yet another aspect, the invention provides a compound of Formula II:

or a pharmaceutically acceptable salt or prodrug thereof, wherein:

Z is —O—, —S—, —C(HR⁷)—, —N(R⁸)— or absent;

R¹ is alkyl, heteroalkyl, cycloalkyl, heterocycloalkyl, aryl,heteroaryl, acyl or cyano;

R² is hydrogen, alkyl, heteroalkyl, alkenyl, alkynyl, nitro,carboxaldehyde, carboxylic acid, ester, amido, cyano, halo or sulfonyl;

R¹⁴ is hydrogen, deuterium or alkyl;

R¹⁵ is hydrogen, hydroxy or amino; or R¹⁴ and R¹⁵ in combination formoxo or methylene;

R¹⁶ and R¹⁷ are independently selected from the group consisting ofhydrogen, halo, alkyl, heteroalkyl and cycloalkyl; or R¹⁶ and R¹⁷ andthe carbon to which they are attached form C₃-C₈ cycloalkyl or C₅-C₈heterocycloalkyl;

R¹⁸ is O or NR¹⁹, wherein R¹⁹ is selected from the group consisting ofhydrogen, alkyl and cyano;

n″ is 1 or 2; and

R⁷ and R⁸ are independently hydrogen, halo, hydroxy, cyano, alkyl oralkoxy.

In yet another aspect, the invention provides a compound of Formula II:

or a pharmaceutically acceptable salt or prodrug thereof, wherein:

Z is —O—, —S—, —S(O)—, —S(O)₂—, —C(O)—, —C(HR⁷)—, —N(R⁸)—, C₁-C₃alkylene, C₁-C₃ heteroalkylene, C₁-C₃ alkenylene or absent;

R¹ is alkyl, heteroalkyl, cycloalkyl, heterocycloalkyl, aryl,heteroaryl, acyl or cyano;

R² is hydrogen, alkyl, heteroalkyl, alkenyl, alkynyl, nitro,carboxaldehyde, carboxylic acid, ester, amido, cyano, halo or sulfonyl;

R¹⁴ is hydrogen, deuterium or alkyl;

R¹⁵ is hydrogen, hydroxy or amino; or R¹⁴ and R¹⁵ in combination formoxo or methylene;

R¹⁶ and R¹⁷ are independently selected from the group consisting ofhydrogen, halo, alkyl, heteroalkyl and cycloalkyl; or R¹⁶ and R¹⁷ andthe carbon to which they are attached form C₃-C₈ cycloalkyl or C₅-C₈heterocycloalkyl;

R¹⁸ is O or NR¹⁹, wherein R¹⁹ is selected from the group consisting ofhydrogen, alkyl and cyano;

n″ is 1 or 2; and

R⁷ and R⁸ are independently hydrogen, halo, hydroxy, cyano, alkyl oralkoxy.

In some embodiments, for a compound of Formula II, R¹ is furtherselected from alkyl, heteroalkyl, cycloalkyl, cycloalkenyl,heterocycloalkyl, aryl, heteroaryl, acyl and cyano.

In some embodiments, R¹ is alkyl, cycloalkyl, heterocycloalkyl, aryl orheteroaryl. In some embodiments, R¹ is cycloalkyl, heterocycloalkyl,aryl or heteroaryl. In some embodiments, R¹ is heterocycloalkyl, aryl orheteroaryl. In some embodiments, R¹ is cycloalkyl, aryl or heteroaryl.In some embodiments, R¹ is aryl or heteroaryl. In a further embodiment,R¹ is phenyl. In another further embodiment, R¹ is pyridyl. In a stillfurther embodiment, the phenyl or pyridyl is substituted with at leastone substituent selected from the group consisting of halo, alkoxy,cyano and alkyl.

In some embodiments, R¹ is selected from the group consisting ofcyclobutyl, cyclohexyl, tetrahydrofuranyl and tetrahydropyranyl.

In some embodiments, R¹ is

wherein each of R^(e) is independently hydrogen or C₁-C₄ alkyl, or twoR^(e)s and the carbon atom to which they are attached form a 4- to8-membered cyclic moiety; each of R^(f) is independently selected fromthe group consisting of halo, alkoxy, cyano and alkyl; and n′″ is 0, 1,2, 3 or 4. In some further embodiments, the 4- to 8-membered cyclicmoiety is an all carbon or heterocyclic ring system.

In some embodiments, R¹ is selected from the group consisting of:

and the rings specified for R¹ may optionally be substituted by one ormore substituents described for aryl and heteroaryl. In a furtherembodiment, the substituent(s) is selected from the group consisting ofhalo, C₁-C₄ alkyl, C₁-C₄ alkoxy and cyano.

In some embodiments, R¹ is cycloalkyl. In other embodiments, R¹ isheterocycloalkyl. In a further embodiment, R¹ is C₃-C₆ cycloalkyl orC₃-C₆ heterocycloalkyl. In yet a further embodiment, R¹ is cyclobutyl.In some embodiments, said cycloalkyl, cyclobutyl or heterocycloalkyl mayoptionally be substituted with one or more substituents described forcycloalkyl or heterocycloalkyl. In a further embodiment, thesubstituent(s) is selected from the group consisting of halo, C₁-C₄alkyl, C₁-C₄ alkoxy and cyano. In another further embodiment, thesubstituent(s) is at least one fluoro.

In some embodiments, R¹ is acyl or cyano. In a further embodiment, R¹ isacetyl.

In some embodiments, R¹ is alkyl. In a further embodiment, the alkyl issubstituted with at least one substituent(s) selected from the groupconsisting of halo, C₁-C₄ alkyl, C₁-C₄ alkoxy and cyano. In anotherfurther embodiment, the alkyl is substituted with at least one fluoro.In some embodiments, R¹ is heteroalkyl.

In some embodiments, R¹ is selected from the group consisting of:

wherein each of the members may optionally be substituted with one ormore substituents selected from the group consisting of cyano, halo,alkyl and alkoxy. In a further embodiment, the substituent(s) isselected from the group consisting of fluoro, C₁-C₄ alkyl, C₁-C₄ alkoxyand cyano.

In some embodiments, R² is nitro, cyano, halo, alkyl, heteroalkyl,alkynyl or alkenyl. In some embodiments, R² is cyano, halo, alkyl,heteroalkyl or alkynyl. In some embodiments, R² is cyano, halo or alkyl.In some embodiments, R² is halo or alkyl. In a further embodiment, R² isfluoroalkyl. In a still further embodiment, R² is C₁-C₄ fluoroalkyl.Exemplary C₁-C₄ fluoroalkyl includes, but is not limited to, —CH₂F,—CHF₂, —CF₂CH₃ and the like.

In some embodiments, R¹⁴ is hydrogen or deuterium. In some embodiments,R¹⁴ is alkyl. In a further embodiment, R¹⁴ is C₁-C₄ alkyl.

In some embodiments, R¹⁵ is hydroxy or amino. In some embodiments, R¹⁵is hydroxy. In some embodiments, R¹⁵ is amino. In a further embodiment,R¹⁵ is NH₂.

In some embodiments, each of R¹⁶ and R¹⁷ is independently hydrogen orfluoro. In some embodiments, each of R¹⁶ and R¹⁷ is hydrogen. In someembodiments, each of R¹⁶ and R¹⁷ is fluoro. In some embodiments, atleast one of R¹⁶ and R¹⁷ is fluoro.

In some embodiments, R¹⁸ is O, N—CN, or NH. In some embodiments, R¹⁸ isO. In some embodiments, R¹⁸ is NH. In some embodiments, R¹⁸ is N—CN.

In some embodiments, R¹⁴ is hydrogen and R¹⁵ is hydroxy or amino. Insome further embodiments, R¹ is aryl or heteroaryl. In a furtherembodiment, R² is cyano, halo or alkyl. In a still further embodiment,R¹⁶ and R¹⁷ are fluoro.

In some embodiments, R¹⁵ is hydroxy or amino and R² is cyano, halo oralkyl. In a further embodiment, R² is fluoroalkyl. In a still furtherembodiment, at least one R¹⁶ and R¹⁷ is fluoro. In a yet still furtherembodiment, n″ is 1.

In some embodiments, R¹⁸ is O or NH and R¹⁴ is hydrogen. In some furtherembodiments, R¹ is aryl or heteroaryl. In a further embodiment, R² iscyano, halo or alkyl. In a still further embodiment, at least one of R¹⁶and R¹⁷ is fluoro.

In some embodiments, n″ is 1. In some further embodiments, R¹⁵ ishydroxy or amino and R¹⁶ and R¹⁷ are fluoro. In a further embodiment, R¹is aryl or heteroaryl. In a still further embodiment, R¹ is phenyl orpyridyl, optionally substituted with one or more substituents selectedfrom the group consisting of halo, alkoxy, cyano and alkyl.

In some embodiments, Z is —O—, —S—, —S(O)—, —S(O)₂—, —S(O₂)N(R⁸)—,—C(O)—, —C(O)O—, —C(HR⁷)—, —N(R⁸)—, —C(O)N(R⁸)—, alkylene, alkenylene,alkynylene, heteroalkylene, or absent. In some embodiments, Z is —O—,—S—, —S(O)—, —S(O)₂—, —C(O)—, —C(HR⁷)—, —N(R⁸)—, C₁-C₃ alkylene, C₁-C₃heteroalkylene, C₁-C₃ alkenylene or absent. In some embodiments, Z is—O—. In other embodiments, Z is —S—. In further embodiments, Z is—C(HR⁷)—. In yet other embodiments, Z is —N(R⁸). In some embodiments, Zis absent.

In another aspect, the invention provides a compound of Formula II-A:

or a pharmaceutically acceptable salt or prodrug thereof, wherein:

Z is —O—, —S—, —C(HR⁷)—, —N(R⁸)— or absent;

R¹ is alkyl, heteroalkyl, cycloalkyl, heterocycloalkyl, aryl,heteroaryl, acyl or cyano;

R² is hydrogen, alkyl, heteroalkyl, alkenyl, alkynyl, nitro,carboxaldehyde, carboxylic acid, ester, amido, cyano, halo or sulfonyl;

R¹⁴ is hydrogen, deuterium or alkyl;

R¹⁵ is hydrogen, hydroxy or amino; or R¹⁴ and R¹⁵ in combination formoxo or methylene;

R¹⁸ is O or NR¹⁹, wherein R¹⁹ is selected from the group consisting ofhydrogen, alkyl and cyano; and

R⁷ and R⁸ are independently hydrogen, halo, hydroxy, cyano, alkyl oralkoxy.

In another aspect, the invention provides a compound of Formula II-A:

or a pharmaceutically acceptable salt or prodrug thereof, wherein:

Z is —O—, —S—, —S(O)—, —S(O)₂—, —C(O)—, —C(HR⁷)—, —N(R⁸)—, C₁-C₃alkylene, C₁-C₃ heteroalkylene, C₁-C₃ alkenylene or absent;

R¹ is alkyl, heteroalkyl, cycloalkyl, heterocycloalkyl, aryl,heteroaryl, acyl or cyano;

R² is hydrogen, alkyl, heteroalkyl, alkenyl, alkynyl, nitro,carboxaldehyde, carboxylic acid, ester, amido, cyano, halo or sulfonyl;

R¹⁴ is hydrogen, deuterium or alkyl;

R¹⁵ is hydrogen, hydroxy or amino; or R¹⁴ and R¹⁵ in combination formoxo or methylene;

R¹⁸ is O or NR¹⁹, wherein R¹⁹ is selected from the group consisting ofhydrogen, alkyl and cyano; and

R⁷ and R⁸ are independently hydrogen, halo, hydroxy, cyano, alkyl oralkoxy.

In some embodiments, for a compound of Formula II-A, R¹ is furtherselected from alkyl, heteroalkyl, cycloalkyl, cycloalkenyl,heterocycloalkyl, aryl, heteroaryl, acyl and cyano.

In some embodiments, R¹ is alkyl, cycloalkyl, heterocycloalkyl, aryl orheteroaryl. In some embodiments, R¹ is cycloalkyl, heterocycloalkyl,aryl or heteroaryl. In some embodiments, R¹ is heterocycloalkyl, aryl orheteroaryl. In some embodiments, R¹ is cycloalkyl, aryl or heteroaryl.In some embodiments, R¹ is aryl or heteroaryl. In a further embodiment,R¹ is phenyl. In another further embodiment, R¹ is pyridyl. In a stillfurther embodiment, the phenyl or pyridyl is substituted with at leastone substituent selected from the group consisting of halo, alkoxy,cyano and alkyl.

In some embodiments, R¹ is selected from the group consisting ofcyclobutyl, cyclohexyl, tetrahydrofuranyl and tetrahydropyranyl.

In some embodiments, R¹ is

wherein each of R^(e) is independently hydrogen or C₁-C₄ alkyl, or twoR^(e)s and the carbon atom to which they are attached form a 4- to8-membered cyclic moiety; each of R^(f) is independently selected fromthe group consisting of halo, alkoxy, cyano and alkyl; and n′″ is 0, 1,2, 3 or 4. In some further embodiments, the 4- to 8-membered cyclicmoiety is an all carbon or heterocyclic ring system.

In some embodiments, R¹ is selected from the group consisting of:

and the rings specified for R¹ may optionally be substituted by one ormore substituents described for aryl and heteroaryl. In a furtherembodiment, the substituent(s) is selected from the group consisting ofhalo, C₁-C₄ alkyl, C₁-C₄ alkoxy and cyano.

In some embodiments, R¹ is cycloalkyl. In other embodiments, R¹ isheterocycloalkyl. In a further embodiment, R¹ is C₃-C₆ cycloalkyl orC₃-C₆ heterocycloalkyl. In yet a further embodiment, R¹ is cyclobutyl.In some embodiments, said cycloalkyl, cyclobutyl or heterocycloalkyl mayoptionally be substituted with one or more substituents described forcycloalkyl or heterocycloalkyl. In a further embodiment, thesubstituent(s) is selected from the group consisting of halo, C₁-C₄alkyl, C₁-C₄ alkoxy and cyano. In another further embodiment, thesubstituent(s) is at least one fluoro.

In some embodiments, R¹ is acyl or cyano. In a further embodiment, R¹ isacetyl.

In some embodiments, R¹ is alkyl. In a further embodiment, the alkyl issubstituted with at least one substituent(s) selected from the groupconsisting of halo, C₁-C₄ alkyl, C₁-C₄ alkoxy and cyano. In anotherfurther embodiment, the alkyl is substituted with at least one fluoro.In some embodiments, R¹ is heteroalkyl.

In some embodiments, R¹ is selected from the group consisting of:

wherein each of the members may optionally be substituted with one ormore substituents selected from the group consisting of cyano, halo,alkyl and alkoxy. In a further embodiment, the substituent(s) isselected from the group consisting of fluoro, C₁-C₄ alkyl, C₁-C₄ alkoxyand cyano.

In some embodiments, R² is nitro, cyano, halo, alkyl, heteroalkyl,alkynyl or alkenyl. In some embodiments, R² is cyano, halo, alkyl,heteroalkyl or alkynyl. In some embodiments, R² is cyano, halo or alkyl.In some embodiments, R² is halo or alkyl. In a further embodiment, R² isfluoroalkyl. In a still further embodiment, R² is C₁-C₄ fluoroalkyl.Exemplary C₁-C₄ fluoroalkyl includes, but is not limited to, —CH₂F,—CHF₂, —CF₂CH₃ and the like.

In some embodiments, R¹⁴ is hydrogen or deuterium. In some embodiments,R¹⁴ is alkyl. In a further embodiment, R¹⁴ is C₁-C₄ alkyl.

In some embodiments, R¹⁵ is hydroxy or amino. In some embodiments, R¹⁵is hydroxy. In some embodiments, R¹⁵ is amino. In a further embodiment,R¹⁵ is NH₂.

In some embodiments, R¹⁸ is O, N—CN, or NH. In some embodiments, R¹⁸ isO. In some embodiments, R¹⁸ is NH. In some embodiments, R¹⁸ is N—CN.

In some embodiments, R¹⁸ is O or NH and R¹⁴ is hydrogen. In some furtherembodiments, R¹ is aryl or heteroaryl. In a further embodiment, R² iscyano, halo or alkyl. In a still further embodiment, at least one of R¹⁶and R¹⁷ is fluoro.

In some embodiments, Z is —O—, —S—, —S(O)—, —S(O)₂—, —S(O₂)N(R⁸)—,—C(O)—, —C(O)O—, —C(HR⁷)—, —N(R⁸)—, —C(O)N(R⁸)—, alkylene, alkenylene,alkynylene, heteroalkylene, or absent. In some embodiments, Z is —O—,—S—, —S(O)—, —S(O)₂—, —C(O)—, —C(HR⁷)—, —N(R⁸)—, C₁-C₃ alkylene, C₁-C₃heteroalkylene, C₁-C₃ alkenylene or absent. In some embodiments, Z is—O—. In other embodiments, Z is —S—. In further embodiments, Z is—C(HR⁷)—. In yet other embodiments, Z is —N(R⁸). In some embodiments, Zis absent.

In still another aspect, the invention provides a compound of FormulaII-B:

or a pharmaceutically acceptable salt or prodrug thereof, wherein:

Z is —O—, —S—, —C(HR⁷)—, —N(R⁸)— or absent;

R¹ is alkyl, heteroalkyl, cycloalkyl, heterocycloalkyl, aryl,heteroaryl, acyl or cyano;

R² is hydrogen, alkyl, heteroalkyl, alkenyl, alkynyl, nitro,carboxaldehyde, carboxylic acid, ester, amido, cyano, halo or sulfonyl;

R¹⁵ is hydroxy or amino;

R¹⁸ is O or NR¹⁹, wherein R¹⁹ is selected from the group consisting ofhydrogen, alkyl and cyano; and

R⁷ and R⁸ are independently hydrogen, halo, hydroxy, cyano, alkyl oralkoxy.

In still another aspect, the invention provides a compound of FormulaII-B:

or a pharmaceutically acceptable salt or prodrug thereof, wherein:

Z is —O—, —S—, —S(O)—, —S(O)₂—, —C(O)—, —C(HR⁷)—, —N(R⁸)—, C₁-C₃alkylene, C₁-C₃ heteroalkylene, C₁-C₃ alkenylene or absent;

R¹ is alkyl, heteroalkyl, cycloalkyl, heterocycloalkyl, aryl,heteroaryl, acyl or cyano;

R² is hydrogen, alkyl, heteroalkyl, alkenyl, alkynyl, nitro,carboxaldehyde, carboxylic acid, ester, amido, cyano, halo or sulfonyl;

R¹⁵ is hydroxy or amino;

R¹⁸ is O or NR¹⁹, wherein R¹⁹ is selected from the group consisting ofhydrogen, alkyl and cyano; and

R⁷ and R⁸ are independently hydrogen, halo, hydroxy, cyano, alkyl oralkoxy.

In some embodiments, for a compound of Formula II-B, R¹ is furtherselected from alkyl, heteroalkyl, cycloalkyl, cycloalkenyl,heterocycloalkyl, aryl, heteroaryl, acyl and cyano.

In some embodiments, R¹ is alkyl, cycloalkyl, heterocycloalkyl, aryl orheteroaryl. In some embodiments, R¹ is cycloalkyl, heterocycloalkyl,aryl or heteroaryl. In some embodiments, R¹ is heterocycloalkyl, aryl orheteroaryl. In some embodiments, R¹ is cycloalkyl, aryl or heteroaryl.In some embodiments, R¹ is aryl or heteroaryl. In a further embodiment,R¹ is phenyl. In another further embodiment, R¹ is pyridyl. In a stillfurther embodiment, the phenyl or pyridyl is substituted with at leastone substituent selected from the group consisting of halo, alkoxy,cyano and alkyl.

In some embodiments, R¹ is selected from the group consisting ofcyclobutyl, cyclohexyl, tetrahydrofuranyl and tetrahydropyranyl.

In some embodiments, R¹ is

wherein each of R^(e) is independently hydrogen or C₁-C₄ alkyl, or twoR^(e)s and the carbon atom to which they are attached form a 4- to8-membered cyclic moiety; each of R^(f) is independently selected fromthe group consisting of halo, alkoxy, cyano and alkyl; and n′″ is 0, 1,2, 3 or 4. In some further embodiments, the 4- to 8-membered cyclicmoiety is an all carbon or heterocyclic ring system.

In some embodiments, R¹ is selected from the group consisting of:

and the rings specified for R¹ may optionally be substituted by one ormore substituents described for aryl and heteroaryl. In a furtherembodiment, the substituent(s) is selected from the group consisting ofhalo, C₁-C₄ alkyl, C₁-C₄ alkoxy and cyano.

In some embodiments, R¹ is cycloalkyl. In other embodiments, R¹ isheterocycloalkyl. In a further embodiment, R¹ is C₃-C₆ cycloalkyl orC₃-C₆ heterocycloalkyl. In yet a further embodiment, R¹ is cyclobutyl.In some embodiments, said cycloalkyl, cyclobutyl or heterocycloalkyl mayoptionally be substituted with one or more substituents described forcycloalkyl or heterocycloalkyl. In a further embodiment, thesubstituent(s) is selected from the group consisting of halo, C₁-C₄alkyl, C₁-C₄ alkoxy and cyano. In another further embodiment, thesubstituent(s) is at least one fluoro.

In some embodiments, R¹ is acyl or cyano. In a further embodiment, R¹ isacetyl.

In some embodiments, R¹ is alkyl. In a further embodiment, the alkyl issubstituted with at least one substituent(s) selected from the groupconsisting of halo, C₁-C₄ alkyl, C₁-C₄ alkoxy and cyano. In anotherfurther embodiment, the alkyl is substituted with at least one fluoro.In some embodiments, R¹ is heteroalkyl.

In some embodiments, R¹ is selected from the group consisting of:

wherein each of the members may optionally be substituted with one ormore substituents selected from the group consisting of cyano, halo,alkyl and alkoxy. In a further embodiment, the substituent(s) isselected from the group consisting of fluoro, C₁-C₄ alkyl, C₁-C₄ alkoxyand cyano.

In some embodiments, R² is nitro, cyano, halo, alkyl, heteroalkyl,alkynyl or alkenyl. In some embodiments, R² is cyano, halo, alkyl,heteroalkyl or alkynyl. In some embodiments, R² is cyano, halo or alkyl.In some embodiments, R² is halo or alkyl. In a further embodiment, R² isfluoroalkyl. In a still further embodiment, R² is C₁-C₄ fluoroalkyl.Exemplary C₁-C₄ fluoroalkyl includes, but is not limited to, —CH₂F,—CHF₂, —CF₂CH₃ and the like.

In some embodiments, R¹⁵ is hydroxy. In some embodiments, R¹⁵ is amino.In a further embodiment, R¹⁵ is NH₂.

In some embodiments, R¹⁸ is O, N—CN, or NH. In some embodiments, R¹⁸ isO. In some embodiments, R¹⁸ is NH. In some embodiments, R¹⁸ is N—CN.

In some embodiments, Z is —O—, —S—, —S(O)—, —S(O)₂—, —S(O₂)N(R⁸)—,—C(O)—, —C(O)O—, —C(HR⁷)—, —N(R⁸)—, —C(O)N(R⁸)—, alkylene, alkenylene,alkynylene, heteroalkylene, or absent. In some embodiments, Z is —O—,—S—, —S(O)—, —S(O)₂—, —C(O)—, —C(HR⁷)—, —N(R⁸)—, C₁-C₃ alkylene, C₁-C₃heteroalkylene, C₁-C₃ alkenylene or absent. In some embodiments, Z is—O—. In other embodiments, Z is —S—. In further embodiments, Z is—C(HR⁷)—. In yet other embodiments, Z is —N(R⁸). In some embodiments, Zis absent.

In some embodiments, a compound of Formula II-B may have an enantiomericexcess of at least about 70%, at least about 71%, at least about 72%, atleast about 73%, at least about 74%, at least about 75%, at least about76%, at least about 77%, at least about 78%, at least about 79%, atleast about 80%, at least about 81%, at least about 82%, at least about83%, at least about 84%, at least about 85%, at least about 86%, atleast about 87%, at least about 88%, at least about 89%, at least about90%, at least about 91%, at least about 92%, at least about 93%, atleast about 94%, at least about 95%, at least about 96%, at least about97%, at least about 98% or at least about 99%. In a further embodiment,the compound has an enantiomeric excess of at least about 90%.

In another aspect, the present disclosure provides a compound orpharmaceutically acceptable salt or prodrug thereof, selected from thegroup consisting of the compounds given in Table 1.

The chemical entities described herein can be synthesized according toone or more illustrative schemes herein and/or techniques known in theart. Materials used herein are either commercially available or preparedby synthetic methods generally known in the art. These schemes are notlimited to the compounds listed in the examples or by any particularsubstituents, which are employed for illustrative purposes. Althoughvarious steps are described and depicted in Schemes 1-27, the steps insome cases may be performed in a different order than the order shown inSchemes 1-27. Various modifications to these synthetic reaction schemesmay be made and will be suggested to one skilled in the art havingreferred to the disclosure contained in this Application.

Unless specified to the contrary, the reactions described herein takeplace at atmospheric pressure, generally within a temperature range from−10° C. to 200° C. Further, except as otherwise specified, reactiontimes and conditions are intended to be approximate, e.g., taking placeat about atmospheric pressure within a temperature range of about −10°C. to about 110° C. over a period of about 1 to about 24 hours;reactions left to run overnight average a period of about 16 hours.

In general, compounds of the invention may be prepared by the followingreaction schemes:

In some embodiments, a compound of Formula 1-9 can be prepared accordingto steps outlined in Scheme 1. The synthesis starts with phenol 1-1.Reaction of 1-1 with chloride 1-2 (wherein R^(g) and R^(h) areindependently alkyl) provides intermediate 1-3. The reaction may becarried out in a suitable organic solvent in the presence of a base.Suitable bases for the reaction include, but are not limited to, organicbases, for example, triethylamine, N,N-diisopropylethylamine,1,4-diazabicyclo[2.2.2]octane, and inorganic bases, for example, sodiumhydroxide, cesium carbonate, cesium bicarbonate, sodium carbonate, andpotassium carbonate. A compound of Formula 1-3 is then subjected to arearrangement reaction to give a compound of Formula 1-4. Elevatedtemperature may be needed for the rearrangement to occur. Thetemperature may be in a range of 100° C. to 300° C. In some embodiments,the temperature is in a range of 180° C. to 240° C. Hydrolysis of acompound of Formula 1-4 provides thiophenol 1-5, which is alkylated toprovide a compound if Formula 1-6. A variety of alkyl groups may beintroduced in Step D. In some embodiments, R^(a) is a C₁-C₄ alkyl. In afurther embodiment, R^(a) is a C₁-C₄ fluoroalkyl. Oxidation of acompound of Formula 1-6 may be accomplished by a variety of methodsknown in the art, including, but not limited to, RuCl₃ catalyzedoxidation in the presence of NaIO₄, oxidation with m-chloroperoxybenzoicacid (mCPBA) and oxidation with Oxone®. Ketone 1-7 is then reduced togive alcohol 1-8, which then undergoes a nucleophilic aromaticsubstitution (SNAr) reaction with a suitable substrate R¹OH to give acompound of Formula 1-9. Temperatures for carrying out the SNAr reactionmay depend on the reactivity of both R¹OH and/or compound 1-8. Thereaction may be carried out in a temperature range from about roomtemperature to 200° C. In some embodiments, the temperature range isfrom room temperature to 60° C. In some other embodiments, thetemperature range is from 60° C. to 100° C. In some other embodiments,the temperature range is from 100° C. to 200° C.

In some other embodiments, a compound of Formula 1-9 can be preparedasymmetrically to give a compound of Formula 2-2 (Scheme 2). Forexample, direct asymmetric reduction of ketone 1-7 (Step A) may beaccomplished chemically or enzymatically. For a recent review onenzymatic reduction of ketones, see Moore, et al. Acc. Chem. Res. 40:1412-1419, 2007. Examples of chemical asymmetric reduction of ketonesinclude, but are not limited to, Corey-Bakshi-Shibata (CBS) reduction,asymmetric hydrogenation and asymmetric transfer hydrogenation. In someembodiments, the asymmetric transfer hydrogenation is catalyzed byruthenium. For examples of methods and catalysts for ruthenium catalyzedtransfer hydrogenation, see U.S. Pat. Nos. 6,184,381 and 6,887,820.Exemplary catalysts for asymmetric transfer hydrogenation include, butare not limited to, the following (shown as the R, R configuration):

The asymmetric transfer hydrogenation may be carried out at or belowroom temperature. In some embodiments, the asymmetric transferhydrogenation is carried out at about 4° C. The alcohol product may havean enantiomeric excess of at least 90%, at least 91%, at least 92%, atleast 93%, at least 94%, at least 95%, at least 96%, at least 97%, oreven higher. It is well understood by one skilled in the art thatchanging the catalyst configuration will lead to a product with theopposite configuration. Chiral alcohol 2-1 can be coupled with asuitable substrate, for example, a phenol, to give a compound of Formula2-2 without significant loss of enantiomeric excess. The loss ofenantiomeric excess (ee) in the coupling step for 2-2 may be less thanabout 1%, less than about 2%, less than about 3%, less than about 4%,less than about 5%, less than about 6% or less than about 8%.

In some embodiments, a compound of Formula 3-6 may be prepared accordingto Scheme 3. The ketone in 1-7 is protected as a ketal to give acompound of Formula 3-1, wherein each of R^(i) and R^(j) isindependently an alkyl group. In addition, R^(i) and R^(j) mayoptionally be connected to form a cyclic ketal. Exemplary structures ofketal 3-1 include, but are not limited to, the following:

A compound of Formula 3-1 and a suitable R¹OH may undergo a nucleophilicaromatic substitution reaction (SNAr) to give biaryl ether 3-2. Asdescribed in Step G of Scheme 1, the reaction temperature of the SNArreaction may depend on the reactivity of the aryl halide (i.e. compound3-1) and/or R¹OH. Ketone 3-3, resulting from the deprotection of ketal3-2, is condensed with an amine to form imine 3-4, wherein R^(k) isalkyl. The imine functional group in a compound of Formula 3-4 may existas a mixture of E and Z isomers. Fluorination of 3-4 can be accomplishedwith a fluorinating reagent, for example,1-(chloromethyl)-4-fluoro-1,4-diazoniabicyclo[2.2.2]octaneditetrafluoroborate, to give difluoroketone 3-5 after acid hydrolysis.Finally, reduction of ketone 3-5 with a hydride donor gives a compoundof Formula 3-6.

A compound of Formula 4-1 (Scheme 4) can be prepared asymmetricallyfollowing the general procedure described above in Scheme 2. In someembodiments, the asymmetric reduction gives a compound of Formula 4-1with an enantiomeric excess of at least about 80%, at least about 81%,at least about 82%, at least about 83%, at least about 84%, at leastabout 85%, at least about 86%, at least about 87%, at least about 88%,at least about 89%, at least about 90%, at least about 91%, at leastabout 92%, at least about 93%, at least about 94%, at least about 95%,at least about 96%, at least about 97%, at least about 98% or evenhigher. The enantiomeric excess of a compound of Formula 2-2 or 4-1 maybe determined by chiral HPLC or Mosher ester analysis. For determinationof ee with Mosher ester, see Hoye, et al. Natural Protocol, 2: 2451,2007.

Alternatively, a compound of Formula 4-1 may be prepared according toScheme 5. Ketone 5-1 is fluorinated to give monofluoroketone 5-2, whichis then converted to silylenol ether, e.g., TBS enol ether 5-3. Othersilyl protecting groups may also be used, for example, triisopropylsilylor diphenyl-t-butylsilyl. The resulting enol ether is furtherfluorinated to give difluoroketone 5-4, which undergoes an asymmetricreduction, such as asymmetric transfer hydrogenation as describedherein, to give chiral alcohol 5-5. Protection of the hydroxy moiety,followed by SNAr reaction and deprotection of the alcohol, provides acompound of Formula 4-1.

Alternatively, a compound of Formula 3-6 can be prepared according toScheme 6. Treatment of aryl halide 3-1 with a hydroxide source givesphenol 6-1. Suitable hydroxide sources include, but are not limited to,sodium hydroxide and potassium hydroxide. Suitable solvents for thereaction include, but are not limited to, DMSO, DMA, DMF and EtOH.Phenol 6-1 can react with a suitable halide via an SNAr reaction to giveether 3-2, which can be converted to a compound of Formula 3-6 asdescribed in Scheme 3.

Compounds of Formulae 7-3 and 7-4 may be prepared according to Scheme 7.For example, condensation of NH₂R^(k) with difluoroketone 7-1, whereinR^(a) is aryl, heteroaryl, alkyl, heteroalkyl, heterocycle, orcycloalkyl, gives intermediate 7-2. In some embodiments, R^(k) is achiral auxiliary. Exemplary chiral auxiliaries include, but are notlimited to:

and enantiomers thereof. Hydride reduction of intermediate 7-2 yields7-3. At this stage, the chiral auxiliary may be cleaved underappropriate conditions, e.g., hydrogenation or acid treatment, to givechiral secondary amine 7-4. In some other embodiments, when a compoundof Formula 7-3 is desirable and R^(k) is not hydrogen, asymmetrichydrogenation or asymmetric transfer hydrogenation is applied onintermediate 7-2 to give a compound of Formula 7-3. For a review onasymmetric hydrogenation and asymmetric transfer hydrogenation, see IwaoOjima ed. Catalytic Asymmetric Synthesis, Wiley-VCH, Inc., 2000, ISBN0-471-29805-0.

In some embodiments, a compound of Formula 8-2 can be prepared accordingto Scheme 8. For example, ketone 3-3 is monofluorinated to give amonofluoroketone of Formula 8-1. The monofluorination can be achievedwith a variety of fluorinating reagents, e.g.,N-Fluoro-O-benzenedisulfonimide, acetyl hypofluorite, Accufluor®,Selectfluor®, Selectfluor® II, or N-fluorobenzenesulfonimide, in thepresence or absence of a base. A compound of Formula 8-1 is reduced togive a compound of Formula 8-2. In some cases, the reduction is highlydiastereoselective to give a compound of Formula 8-2 with greater than80%, greater than 82%, greater than 84%, greater than 86%, greater than88%, greater than 90%, greater than 92%, greater than 94%, greater than96% or even greater than 96% diastereoselectivity. In some cases, thereduction is highly enantioselective to give a compound of Formula 8-2with greater than 80%, greater than 82%, greater than 84%, greater than86%, greater than 88%, greater than 90%, greater than 92%, greater than94%, greater than 96% or even greater than 96% enantioselectivity.Reduction conditions to achieve high enantioselectivity include, but arenot limited to, asymmetric transfer hydrogenation and enzymaticreduction as described herein.

In some embodiments, a compound of Formula 9-6 may be prepared accordingto Scheme 9, wherein R¹³ is hydrogen, alkyl or fluoro. The hydroxy groupof compound 9-1 may be protected, for example, with an acyl ormethoxymethyl ether (MOM) group, to give a compound of Formula 9-2.Benzylic bromination in Step B may be carried out with a bromide source,e.g., N-bromosuccinimide, in the presence of a radical initiator, e.g.,2,2′-azobis(2-methylpropionitrile) (AIBN) or benzoyl peroxide. Thebromide of compound 9-3 can be replaced with a hydroxy group in asolvent comprising water in the presence of a silver salt, e.g., Ag₂CO₃or AgClO₄ or AgBF₄. Finally, fluorination of a compound of Formula 9-4followed by deprotection gives a compound of Formula 9-6. In some cases,direct benzylic oxidation may be used for converting a compound ofFormula 9-2 to a compound of Formula 9-4, thus bypassing an intermediatebromination step.

In some embodiments, a compound of Formula 10-7 can be preparedaccording to Scheme 10. For example, a compound of Formula 10-3 may beprepared from a compound of Formula 3-2 following a similar sequence asoutlined in Scheme 9. Further functional group manipulations lead to acompound of Formula 10-7.

Alternatively, a compound of Formula 10-3 can be deprotected to givediketone 11-1, which is fluorinated to give difluorodiketone 11-2.Asymmetric reduction of 11-2 provides diol 11-3. In some embodiments, afluorination step is performed to give a compound of Formula 10-7.

Alternatively, a compound of Formula 10-7 may be prepared according toScheme 12. For example, difluoroketone 12-2 is reduced to givehydroxyketone 12-3. The reduction maybe enantioselective under transferhydrogenation conditions with a Ru-catalysis as described herein. One ofthe aryl fluorines may be selectively displaced with an alkyl thiol togive a compound of Formula 12-4. Oxidation, fluorination, nucleophilicaromatic substitution (SNAr) and asymmetric reduction give a compound ofFormula 10-7.

In some embodiments, a compound of Formula 13-4 can be preparedaccording to Scheme 13. An aryl sulfide of Formula 13-1 is treated withH₂N—R^(b) and an oxidant, e.g., diacetoxyiodobenzene ordipivaloyloxyiodobenzene, in a suitable solvent, such as acetonitrile,to obtain aryl sulfinimide 13-2. In some embodiments, wherein R^(a) isfluoroalkyl, the addition of rhodium(II) acetate or Rh₂(esp)₂ catalystalong with magnesium oxide is helpful in Step A. Oxidation of arylsulfinimide 13-2 to substituted sulfoximine 13-3 may be accomplishedwith catalytic ruthenium(III) chloride and sodium periodate in asuitable solvent, such as a mixture of water, acetonitrile, and carbontetrachloride. Substituted sulfoximine 13-3 is then manipulatedsimilarly as described in Schemes 3 and 4 to afford sulfoximines ofFormula 13-4 as a diastereomeric mixture. The diastereomers may beseparated by column chromatography.

In some embodiments, a compound of Formula 14-10 can be preparedaccording to steps outlined in Scheme 14, wherein R¹ is alkyl,heteroalkyl, cycloalkyl, heterocycloalkyl, aryl or heteroaryl; R² ishalo, cyano, alkyl, alkenyl or alkynyl; and R¹⁶ and R¹⁷ are fluoro oralkyl, or R¹⁶ and R¹⁷ and the carbon to which they are attached formC₃-C₈ cycloalkyl or C₅-C₈ heterocycloalkyl. The synthesis commences withcompounds of Formula 14-1. Orthoiodination of 14-1 provides compound14-2. The reaction may be carried out in a suitable organic solvent inthe presence of iodine and a palladium catalyst at an elevatedtemperature, if needed. After esterification of 14-2, the resultingester 14-3 may undergo a transition-metal catalyzed coupling reactionwith a thioate, e.g., potassium ethanethioate or sodium ethanethioate,to give compounds of Formula 14-4. Suitable transition-metal catalystsinclude, but are not limited to, Pd(PPh₃)₄, Pd₂(dba)₃ chloroform complexor Pd(OAc)₂, in the presence or absence of a suitable ligand. Hydrolysisof a compound of Formula 14-4 followed by alkylation of the resultingthiophenol intermediate with an alkyl halide, e.g., methyl iodide, givesa compound of Formula 14-5. The hydrolysis and alkylation may be carriedout in a one-pot procedure without purification. In some embodiments,this is carried out by treating a compound of Formula 14-4 with acarbonate base in a suitable solvent at or near room temperature for aperiod ranging from 0.1 to 24 hours, followed by addition of an alkylhalide. Carbonate bases include, but are not limited to, sodiumcarbonate, potassium carbonate, cesium carbonate, potassium bicarbonateand cesium bicarbonate. Oxidation of a compound of Formula 14-5 to givea compound of Formula 14-6 may be accomplished by a variety of methodsknown in the art, including, but not limited to, RuCl₃ catalyzedoxidation in the presence of NaIO₄, oxidation with m-chloroperoxybenzoicacid (mCPBA), and oxidation with Oxone®. A compound of Formula 14-6 isthen subjected to a nucleophilic aromatic substitution (SNAr) reactionwith R¹OH (wherein R¹ is alkyl, aryl or heteroaryl) to give a compoundof Formula 14-7. Temperature for carrying out the SNAr reaction maydepend on the reactivity of both R¹OH and/or a compound of Formula 14-6.The reaction may be carried out at a temperature ranging from −10° C. to200° C. In some embodiments, the temperature range is from 30° C. to120° C. In some other embodiments, the temperature range is from 0° C.to room temperature. Cyclization of a compound of Formula 14-7 may beeffected with a base, e.g., sodium hydride, in a suitable solvent toyield a compound of Formula 14-8. After the cyclization, a variety ofR¹⁶ and R¹⁷ groups may be introduced. In some embodiments, a compound ofFormula 14-8 is difluorinated to give a compound of Formula 14-9, formedby treatment with a fluorinating agent, e.g.,1-(chloromethyl)-4-fluoro-1,4-diazo niabicyclo[2.2.2]octaneditetrafluoroborate (Selectfluor®), in the presence of suitable base,e.g., sodium carbonate. Reduction of a compound of Formula 14-9 yields acompound of Formula 14-10. In some embodiments, the reduction is carriedout with a hydride, e.g., sodium borohydride and sodiumtriacetoxyborohydride, to give a racemic mixture. In some embodiments,an asymmetric reduction is carried out as described above (see Scheme 2)to give an enantiomer having an enantiomeric excess as disclosed herein.

Alternatively, a compound of Formula 14-8 may be prepared according toScheme 15. For example, lithiation of a compound of Formula 15-1followed by trapping of the resulting lithio intermediate with asuitable electrophile gives a compound of Formula 15-2. In someembodiments, the electrophile is N,N-dimethylformamide and R² is —CHO.In a further embodiment, —CHO is converted to —CHF₂ through addition ofa fluorinating reagent, e.g., diethylaminosulfur trifluoride. One of thefluorines in a compound of Formula 15-2 may be selectively displacedwith a thiomethoxide, e.g., sodium thiomethoxide, to give compounds ofFormula 15-3. The reaction temperature may be in a range of −50 to 40°C. In some embodiments, the temperature is at or about 0° C. Oxidationof a compound of Formula 15-3, followed by SNAr reaction with R¹OH andbase-mediated cyclization provides a compound of Formula 14-8.

In some embodiments, a compound of Formula 16-6 may be preparedaccording to Scheme 16. Oxidation of a compound of Formula 15-3 gives acompound of Formula 16-1. The oxidation may be accomplished with Oxone®or mCPBA. The amount of oxidant used for the oxidation may be about 1.5equivalent, about 1.4 equivalent, about 1.3 equivalent, about 1.2equivalent, about 1.1 equivalent or about 1.0 equivalent. SNAr reactionof a compound of Formula 16-1 with R¹OH (wherein R¹ is alkyl,heteroalkyl, cycloalkyl, heterocycloalkyl, aryl or heteroaryl) in thepresence of a base gives a compound of Formula 16-2. At this stage, asulfoximine moiety may be installed to give a compound of Formula 16-3through a transition-metal catalyzed insertion of a suitable nitrogendonor. Suitable transition-metal catalysts include, but are not limitedto, copper and rhodium catalysts, e.g.,bis(rhodium(α,α,α′,α′-tetramethyl-1,3-benezenedipropionic acid)) anddirhodium tetraacetate. Suitable nitrogen donors include, but are notlimited to, PhI=NNs, cyanamide, and fluoroalkylamides, e.g.,trifluoromethyl acetamide. Cyclization of a compound of Formula 16-3 togive a compound of Formula 16-4 may be achieved with a base, e.g.,sodium hydride, at about room temperature. Finally, reduction of acompound of Formula 16-5 as outlined in Scheme 14 provides a compound ofFormula 16-6. A compound of Formula 16-6 may exist as a mixture ofdiastereomers and/or enantiomers. Diastereomers may be separated byconventional column chromatography, while enantiomers may be separatedby chiral column chromatography.

In some embodiments, a compound of Formula 17-5 can be preparedaccording to Scheme 17. Reaction of a compound of Formula 15-3 withcyanamide in the presence of an oxidant, e.g., (diacetoxyiodo)benzene,affords a compound of Formula 15-1. Further oxidation of a compound ofFormula 15-1 provides a compound of Formula 15-2, which undergoes SNArreaction, cyclization and reduction to give a compound of Formula 17-5.

In some embodiments, compounds of Formula 18-8a and 18-8b may beprepared according to Scheme 18. For example, pyridine 18-1 may beconverted to alkylaryl derivative 18-2 in Step A, wherein R⁴ is, forexample, trifluoromethyl. The ketone may be converted to protected enolether 18-3, then fluorinated to give fluoroketal 18-4. Treatment of acompound of Formula 18-4 with a suitable hydroxide source as describedin Scheme 6 gives a mixture of phenols 18-5a and 18-5b. The phenols canundergo an SNAr reaction with a suitable halide to give aryl ethers ofFormulae 18-6a and 18-6b, which may be deprotected to give the resultantketones. In some embodiments, a compound of Formula 18-7 is reduced witha hydride source to give a racemic mixture. In other embodiments, anasymmetric reduction is carried out as described in Scheme 2, affordingalcohols 18-8a and 18-8b, separable by methods known to one skilled inthe art, such as, for example, conventional column chromatography.

Alternatively, a compound of Formula 18-8a can be prepared according toScheme 19. For example, conversion of a compound of Formula 19-1 to aphenol is followed by coupling to form an aryl ether and deprotection togive a ketone of Formula 18-7b as described above. Ketone 18-7b isconverted to silyl enol ether 19-5 using a suitable silyl protectinggroup, including, for example, tert-butyldimethylsilyl,triisopropylsilyl or diphenyl-t-butylsilyl. The resulting silyl enolether is fluorinated to give fluoroketone 18-7a, which undergoes anasymmetric reduction, such as asymmetric transfer hydrogenation asdescribed herein, to give chiral alcohol 18-8a.

A compound of Formula 20-9 can be prepared following the generalprocedure outlined in Scheme 20, wherein the aryl fluoride of a compoundof Formula 20-1 is displaced with an alkyl thiol to give 20-2. Formationof benzaldehyde 20-3 may be followed by, for example, a Wittig reaction,to give alkene 20-4, which is reduced to an alkane of Formula 20-5 undersuitable conditions. In some embodiments, a Dieckmann condensationreaction is followed by a decarboxylation to give ketone 20-6. Oxidationof a compound of Formula 20-6 to give a compound of Formula 20-7 may beaccomplished by a variety of methods known in the art, including, butnot limited to, RuCl₃ catalyzed oxidation in the presence of NaIO₄,oxidation with m-chloroperoxybenzoic acid (mCPBA) and oxidation withOxone®. Protection of the ketone, for example, as the cyclic ketal(20-8) is followed by the general procedure outlined in Scheme 19 togive a compound of Formula 20-9.

In some embodiments, a compound of Formula 21-7 may be preparedaccording to Scheme 21. Formation of a chloropyridine of Formula 21-2may be followed by an SNAr reaction with a suitable alcohol of formulaR¹OH as described above to give a compound of Formula 21-3. Aryl bromide21-3 may undergo a transition-metal catalyzed coupling reaction with athioate to give a compound of Formula 21-4, analogously to the proceduredetailed in Scheme 14. Hydrolysis, alkylation with a suitable alkylhalide and oxidation affords a compound of Formula 21-7.

In some embodiments, a ketone of Formula 22-1 may be reduced to give22-2, optionally with high enantioselectivity using asymmetric transferhydrogenation or enzymatic reduction conditions as described herein. Acompound of Formula 22-2 and a suitable coupling partner, including, butnot limited to, a boronic acid of formula R¹B(OH)₂, may undergo acoupling reaction to give a compound of Formula 22-3.

In some embodiments, R¹ can be coupled to a compound of Formula 23-1 or23-4 via a reaction scheme represented generally in Scheme 23. In someembodiments, wherein Z is —N(R⁸), an aryl halide of Formula 23-1 iscoupled to a suitably substituted amine, i.e. NHR¹R⁸, via aBuchwald-Hartwig amination to give a compound of Formula 23-2. In afurther embodiment, Step A is a cross coupling reaction, including, butnot limited to, a Stille, Negishi or Suzuki reaction, wherein an arylhalide of Formula 23-1 is combined with an appropriate reactantcontaining R¹ and a suitable catalyst to afford a compound of Formula23-3. In other embodiments, a compound of Formula 23-4 undergoes an SNArreaction and a subsequent deprotection to give a compound of Formula23-3. R¹ in a compound of Formula 23-5 may be, for example, morpholine,wherein a C—N bond connects said morpholine to the aryl ring. In stillother embodiments, Z is —S—, and R¹S— is attached to a compound ofFormula 23-1 via an SNAr reaction to give a compound of Formula 23-6.

In some additional embodiments, a compound of Formula 24-2 can beprepared according to steps outlined in Scheme 24. A compound of Formula12-6 is prepared via an asymmetric variation of the sequence presentedin Scheme 12, then reacted with an amine, i.e. (3S)-3-piperidinolhydrochloride, to give a compound of Formula 24-1. Asymmetric reductionas described above gives a compound of Formula 24-2.

In some embodiments, a compound of Formula 25-3 or 25-4 may be preparedaccording to steps outlined in Scheme 25. For example, 25-2 may beprepared via a cross coupling reaction, i.e., a Suzuki reaction with aboronic acid of formula R¹B(OH)₂. Hydride reduction of 25-2 gives amixture of alcohols 25-3 and unsaturated alcohol 25-4.

In some embodiments, a compound of Formula 26-5 can be synthesizedaccording to Scheme 26. For example, treatment of compound 26-1 withbase can provides difluoroalkene 26-2. The amount of base may be about 1equivalent to avoid byproduct formation. The olefin in 26-2 may behydrogenated to give a compound of Formula 26-3 with cis difluoroconfiguration. At this stage, an alkoxy group can be introduced bydisplacement of the aryl fluoride in 26-3 by an alcohol in the presenceof a base. Finally, deprotection followed by reduction of the resultingketone provides a compound of Formula 26-5. Intermediate 26-3 may beseparated, for example, by chiral column chromatography, to give bothenantiomers, each of which can be functionalized as outlined in Scheme26 to provide either enantiomer of a compound of Formula 26-5.

In some embodiments, a compound of Formula 27-6 may be synthesizedaccording to Scheme 27. For example, alkylation of compound 27-1provides a compound of Formula 27-2, wherein R¹ is alkyl, cycloalkyl, orheterocycloalkyl. A compound of Formula 27-2 can be selectivelyiodinated to give aryl iodide 27-3. A compound of Formula 27-3 can bedifluorinated as previously described. Installation of a CF₃ groupfollowed by asymmetric reduction provides a compound of Formula 27-6.

In some other embodiments, a compound of a formula given in Table 1 issynthesized according to one of the general routes outlined in Schemes1-27, Examples 1-35 or by methods generally known in the art.

TABLE 1 No. Structure m/z ¹H NMR Data 1

393 (M + H) (400 MHz, CDCl₃): δ 7.81 (d, 1H), 7.00-6.89 (m, 3H),6.73-6.71 (m, 1H), 6.35 (t, 1H), 5.66-5.65 (m, 1H), 3.19- 3.13 (m, 2H),2.96-2.90 (m, 1H), 2.50-2.40 (m, 1H), 2.30-2.24 (m, 1H) 2

377 (M + H) (400 MHz, CDCl₃): δ 7.81 (d, 1H), 6.96 (d, 1H), 6.73-6.68(m, 1H), 6.62- 6.61 (m, 2H), 6.36 (t, 1H), 5.66-5.65 (m, 1H), 3.22-3.10(m, 2H), 2.96-2.90 (m, 1H), 2.50-2.40 (m, 1H), 2.29-2.24 (m, 1H) 3

376, 378 (M + H) (400 MHz, CDCl₃): δ 8.49 (s, 1H), 8.36 (s, 1H), 7.81(d, 1H), 7.44-7.43 (m, 1H), 6.89 (d, 1H), 6.36 (t, 1H), 5.67-5.66 (m,1H), 3.23-3.16 (m, 2H), 2.99-2.92 (m, 1H), 2.51-2.42 (m, 1H), 2.32-2.25(m, 1H) 4

367 (M + H) (400 MHz, CDCl₃): δ 8.76 (s, 1H), 8.66 (s, 1H), 7.86 (d,1H), 7.65-7.64 (m, 1H), 6.93 (d, 1H), 6.38 (t, 1H), 5.71-5.65 (m, 1H),3.20-3.16 (m, 2H), 2.96-2.90 (m, 1H), 2.50-2.42 (m, 1H), 2.37-2.24 (m,1H) 5

360 (M + H) (400 MHz, CDCl₃): δ 8.41 (s, 1H), 8.32 (s, 1H), 7.82 (d,1H), 7.22-7.17 (m, 1H), 6.92 (d, 1H), 6.37 (t, 1H), 5.70-5.60 (m, 1H),3.23-3.18 (m, 2H), 2.99-2.97 (m, 1H), 2.54-2.40 (m, 1H), 2.34-2.22 (m,1H) 6

433 (M − H + 46) (400 MHz, CDCl₃): δ 7.77 (d, 1H), 6.91 (d, 1H),6.54-6.50 (m, 1H), 6.42- 6.38 (m, 2H), 6.39 (t, 1H), 5.67-5.63 (m, 1H),3.80 (s, 3H), 3.23-3.15 (m, 2H), 2.99-2.92 (m, 1H), 2.50-2.45 (m, 1H),2.30-2.23 (m, 1H) 7

7a, 429, 431 (M + Na) 7b, 429, 431 (M + Na) 7a: (400 MHz, CDCl₃): δ 7.81(d, 1H), 7.01-6.98 (m, 1H), 6.91-6.89 (m, 2H), 6.75-6.71 (m, 1H), 6.34(t, 1H), 5.58-5.53 (m, 1H), 3.48-3.40 (m, 1H), 3.22 (d, 1H), 2.66-2.59(m, 1), 1.98- 1.93 (m, 1H), 1.46 (d, 3H) 7b: (400 MHz, CDCl₃): δ 7.81(d, 1H), 7.01-6.97 (m, 1H), 6.92 (d, 1H), 6.89-6.88 (m, 1H), 6.73-6.69(m, 1H), 6.38 (t, 1H), 5.70-5.67 (m, 1H), 3.71- 3.64 (m, 1H), 3.25 (d,1H), 2.47-2.41 (m, 1H), 2.14-2.06 (m, 1H), 1.36 (d, 3H) 8

429, 431 (M + H) (400 MHz, CDCl₃): δ 7.90 (d, 1H), 7.06-7.03 (m, 1H),6.98 (d, 1H), 6.94- 6.92 (m, 1H), 6.78-6.74 (m, 1H), 6.42 (t, 1H), 5.50(d, 1H), 3.61-3.43 (m, 2H), 3.24 (s, 1H) 9

413 (M + H) (400 MHz, CDCl₃): δ 7.90 (d, 1H), 7.01 (d, 1H), 6.80-6.73(m, 1H), 6.70- 6.63 (m, 2H), 6.43 (t, 1H), 5.50 (m, 1H), 3.60-3.43 (m,2H), 3.30 (d, 1H) 10

375, 377 (M − OH) (400 MHz, CDCl₃): δ 7.85 (d, 1H), 7.04-7.00 (m, 1H),6.97-6.95 (m, 1H), 6.84-6.77 (m, 2H), 6.18 (t, 1H), 5.58- 5.53 (m, 1H),3.59-3.50 (m, 1H), 3.34-3.26 (m, 1H), 2.60-2.50 (m, 1H), 2.31 (d, 1H),2.21-2.13 (m, 1H) 11

437 (M + NH₄) (400 MHz, CDCl₃): δ 7.94 (d, 1H), 7.33-7.29 (m, 1H),7.23-7.21 (m, 1H), 7.13-7.09 (m, 1H), 7.00 (d, 1H), 6.43 (t, 1H), 5.51(d, 1H), 3.60-3.43 (m, 2H), 3.30 (br s, 1H) 12

451 (M − H) (400 MHz, CDCl₃): δ 8.01 (d, 1H), 6.97 (d, 1H), 6.82-6.55(m, 4H), 5.76- 5.64 (m, 1H), 5.35-5.26 (m, 2H), 3.54-3.44 (m, 2H),3.31-3.18 (m, 1H), 3.06-2.96 (m, 2H) 13

451 (M + NH₄) (400 MHz, CDCl₃): δ 8.0 (d, 1H), 7.33-7.30 (m, 1H),7.23-7.21 (m, 1H), 7.13-7.09 (m, 1H), 6.92 (d, 1H), 6.62 (m, 1H),3.58-3.49 (m, 2H), 3.34-3.20 (m, 1H), 1.84-1.82 (m, 3H) 14

364 (M − H) (400 MHz, CDCl₃): δ 7.9 (d, 1H), 6.97 (d, 1H), 6.73-6.67 (m1H), 6.64- 6.58 (m, 1H), 5.83-5.79 (m, 1H), 6.57-6.53 (m, 1H), 4.22 (d,1H), 3.20- 3.10 (m, 1H), 2.95-2.85 (m, 2H), 2.60-2.50 (m, 1H), 2.25-2.16(m, 1H) 15

420 (M + H) (400 MHz, CDCl₃): δ 7.94 (d, 1H), 7.33-6.98 (m, 4H), 6.44(t, 1H), 5.51 (d, 1H), 3.61-3.45 (m, 2H) 16

437/439 (M − H + 46) (400 MHz, CDCl₃): δ 7.81 (d, 1H), 7.00-6.98 (m,1H), 6.94 (d, 1H), 6.89- 6.88 (m, 1H), 6.74-6.71 (m, 1H), 6.35 (t, 1H),5.67-5.65 (m, 1H), 3.21-3.13 (m, 2H), 2.96-2.89 (m, 1H), 2.50-2.41 (m,1H), 2.30-2.23 (m, 1H) 17

393 (M + H) 18

391, 393 (M + H) (400 MHz, CDCl₃): δ 8.15 (d, 1H), 7.14 (d, 1H), 7.12(t, 1H), 7.07-7.04 (m, 1H), 6.96-6.93 (m, 1H), 6.80-6.76 (m, 1H),3.23-3.20 (m, 2H), 2.90-2.87 (m, 2H) 19

376 (M + H) (400 MHz, CDCl₃): δ 7.81 (d, 1H), 6.92 (d, 1H), 6.72-6.67(m, 1H), 6.62 (t, 1H), 6.63-6.59 (m, 2H), 4.96-4.94 (m, 1H), 3.18-3.10(m, 1H), 2.99-2.92 (m, 1H), 2.51-2.41 (m, 1H), 2.30-2.00 (m, 3H) 20

361 (M + H) (400 MHz, CDCl₃): δ 7.76 (d, 1H), 6.87 (d, 1H), 6.69-6.63(m, 1H), 6.60- 6.55 (m, 2H), 6.18 (t, 1H), 3.37 (t, 2H), 2.93 (t, 2H),2.20-2.17 (m, 2H) 21

359 (M + H) (400 MHz, CDCl₃): δ 7.88 (d, 1H), 7.47-7.45 (m, 1H),6.93-6.90 (m, 2H), 6.71-6.60 (m, 3H), 6.22 (t, 1H), 3.49- 3.48 (m, 1H)22

421 (M − H + 46) (400 MHz, CDCl₃): δ 7.79 (d, 1H), 6.90 (d, 1H),6.72-6.66 (m, 1H), 6.64- 6.57 (m, 2H), 6.19 (t, 1H), 4.85-4.81 (m, 1H),3.60-3.44 (m, 3H), 3.21-2.99 (m, 2H) 23

437 (M − H + 46) (400 MHz, CDCl₃): δ 7.83 (d, 1H), 6.98 (d, 1H),6.74-6.69 (m, 1H), 6.64- 6.62 (m, 2H), 6.36 (t, 1H), 5.37 (brs, 1H),4.65-4.63 (m, 1H), 3.45-3.39 (m, 2H), 2.92-2.88 (m, 1H) 24

391 (M + H) (400 MHz, CDCl₃): δ 7.77 (d, 1H), 6.90 (d, 1H), 6.71-6.65(m, 1H), 6.62- 6.36 (m, 2H), 6.23 (t, 1H), 3.94-3.71 (m, 3H), 2.97-2.89(m, 2H), 2.84 (s, 1H), 2.40-2.22 (m, 2H) 25

410, 412 (M + H) (400 MHz, CDCl₃) δ 8.55-8.54 (m, 1H), 8.40-8.39 (m,1H), 7.91 (d, 1H), 7.52-7.49 (m, 1H), 6.93 (d, 1H), 6.44 (t, 1H),5.53-5.49 (m, 1H), 3.64-3.48 (m, 2H), 3.35 (d, 1H) 26

473, 475 (M − H + 46) (400 MHz, CDCl₃): δ 7.86 (d, 1H), 7.26-7.22 (m,2H), 7.05-6.95 (m, 1H), 6.86 (d, 1H), 6.41 (t, 1H), 5.51-5.47 (m, 1H),3.58-3.51 (m, 2H), 3.26 (brd s, 1H) 27

403 (M + H) (400 MHz, CDCl₃): δ 8.82 (s, 1H), 8.71 (s, 1H), 7.95 (d,1H), 7.73-7.71 (m, 1H), 6.95 (d, 1H), 6.44 (t, 1H), 5.55-5.50 (m, 1H),3.60-3.51 (m, 2H), 3.29 (d, 1H) 28

412, 414 (M − H) (400 MHz, CDCl₃): δ 8.27 (d, 1H), 7.17-7.14 (m, 1H),7.03-7.02 (m, 1H), 6.97 (d, 1H), 6.88-6.85 (m, 1H) 29

403/405 (M − H) (400 MHz, CDCl₃): δ 8.31 (d, 1H), 7.44-7.41 (m, 1H),7.32-7.31 (m, 1H), 7.24-7.20 (m, 1H), 6.97 (d, 1H) 30

385, 387 (M − H) (400 MHz, CDCl₃): δ 8.28 (d, 1H), 7.42-7.40 (m, 1H),7.31-7.26 (m, 1H), 7.22-7.19 (m, 1H), 6.97 (d, 1H), 6.45 (t, 1H) 31

[M − H] 374 (400 MHz, CDCl₃): δ 8.14 (d, 1H), 7.67-7.61 (m, 2H),7.48-7.47 (m, 1H), 6.80 (d, 1H), 6.24 (t, 1H), 2.98 (s, 3H) 32

[M − 1] 374 (400 MHz, CDCl₃): δ 8.15 (d, 1H), 7.12-7.08 (m, 1H),7.01-6.99 (m, 1H), 6.89 (d, 1H), 6.85-6.81 (m, 1H), 6.24 (t, 1H), 2.97(s, 3H) 33

[M − H] 365 (400 MHz, CDCl₃): δ 8.20 (d, 1H), 7.39-7.35 (m, 1H),7.29-7.27 (m, 1H), 7.20-7.16 (m, 1H), 6.90 (d, 1H), 6.26 (t, 1H), 2.90(s, 3H) 34

[M − H] 358 (400 MHz, CDCl₃): δ 8.15 (d, 1H), 6.91 (d, 1H), 6.85-6.79(m, 1H), 6.77- 6.70 (m, 2H), 6.24 (t, 1H), 2.97 (s, 3H) 35

[M − H] 391 (400 MHz, CDCl₃): δ 8.07 (d, 1H), 7.13-7.05 (m, 1H),7.04-6.95 (m, 2H), 6.89-6.85 (m, 1H), 6.26 (t, 1H), 5.60 (s, 2H) 36

[M + H] 419 (400 MHz, CDCl₃): δ 8.29 (d, 1H), 7.13-7.09 (m, 1H), 7.02(t, 1H), 7.01- 6.99 (m, 1H), 6.96 (d, 1H), 6.86-6.82 (m, 1H), 4.11 (s,2H), 2.34 (s, 6H) 37

[M − H] 358 (400 MHz, CDCl₃): δ 8.07 (d, 1H), 7.07-7.03 (m, 1H), 7.01(d, 1H), 6.96- 6.94 (m, 1H), 6.81-6.77 (m, 1H), 6.15 (t, 1H), 2.68 (s,3H) 38

[M − H + 18] 413 (400 MHz, CDCl₃): δ 8.03 (d, 1H), 7.24 (d, 1H),7.12-7.08 (m, 1H), 6.96- 6.94 (m, 1H), 6.81-6.77 (m, 1H), 6.41 (t, 1H)39

[M − H + 18] 404 (400 MHz, CDCl₃): δ 8.09 (d, 1H), 7.37-7.34 (m, 1H),7.29 (d, 1H), 7.22- 7.21 (m, 1H), 7.14-7.10 (m, 1H), 6.43 (t, 1H) 40

[M − H + 18] 386 (400 MHz, CDCl₃): δ 8.03 (d, 1H), 7.68-7.62 (m, 2H),7.45-7.43 (m, 1H), 7.40-7.36 (m, 1H), 7.17 (d, 1H), 6.41 (t, 1H) 41

[M − H + 46] 445 (400 MHz, CDCl₃): δ 8.01 (d, 1H), 7.06 (d, 1H),7.04-7.01 (m, 1H), 6.91- 6.88 (m, 1H), 6.76-6.71 (m, 1H), 6.47 (t, 1H),5.21 (d, 2H), 2.69 (t, 1H) 42

[M − H + 46] 436 (400 MHz, CDCl₃): δ 8.06 (d, 1H), 7.28-7.25 (m, 1H),7.15-7.12 (m, 2H), 7.07-7.03 (m, 1H), 6.50 (t, 1H), 5.21 (d, 2H), 2.70(t, 1H) 43

[M − H + 46] 418 (400 MHz, CDCl₃): δ 8.00 (d, 1H), 7.59-7.56 (m, 1H),7.38-7.37 (m, 1H), 7.36-7.31 (m, 1H), 7.00 (d, 1H), 6.48 (t, 1H), 5.22(d, 2H), 2.71 (t, 1H) 44

[M − H + 46] 445 (400 MHz, CDCl₃): δ 8.01 (d, 1H), 7.06 (d, 1H),7.04-7.01 (m, 1H), 6.91- 6.88 (m, 1H), 6.76-6.71 (m, 1H), 6.47 (t, 1H),5.21 (d, 2H), 2.69 (t, 1H) 45

[M + H] 451 (400 MHz, CDCl₃): δ 8.06 (d, 1H), 7.67 (s, 1H), 7.12-7.03(m, 4H), 6.93 (s, 1H), 6.77 (br d, 1H), 5.92 (t, 1H), 5.76 (d, 2H) 46

[M − H + 46] 463 (400 MHz, CDCl₃): δ 8.03 (d, 1H), 7.07 (d, 1H),7.06-7.03 (m, 1H), 6.93- 6.91 (m, 1H), 6.78-6.74 (m, 1H), 6.42 (t, 1H),5.26 (d, 2H) 47

[M + H] 482 (400 MHz, CDCl₃): δ 8.04 (d, 1H), 7.05-7.01 (m, 2H),6.90-6.89 (m, 1H), 6.75-6.71 (m, 1H), 6.67 (s, 1H), 6.43 (t, 1H), 4.50(br s, 2H) 3.40-3.30 (m, 2H) 48

[M + H] 484 (400 MHz, CDCl₃): δ 8.04 (d, 1H), 7.07 (t, 2H), 7.03-6.99(m, 2H), 6.89- 6.87 (m, 1H), 6.74-6.70 (m, 1H), 4.40 (s, 2H), 4.05-3.98(m, 2H), 3.48-3.40 (m, 2H), 2.89-2.81 (m, 1H), 1.97-1.90 (m, 2H),1.52-1.41 (m, 3H) 49

[M + H] 484 (400 MHz, CDCl₃): δ 8.04 (d, 1H), 7.09 (t, 2H), 7.03-7.00(m, 2H), 6.89- 6.88 (m, 1H), 6.74-6.70 (m, 1H), 4.40 (s, 2H), 3.94-3.89(m, 1H), 3.78-3.71 (m, 1H), 3.56-3.49 (m, 1H), 3.39-3.33 (m, 1H),2.86-2.79 (m, 1H), 2.06-1.97 (m, 1H), 1.80-1.72 (m, 1H), 1.66-1.46 (m,3H) 50

[M − H] 338 (400 MHz, CDCl₃): δ 8.20 (d, 1H), 7.08-7.04 (m, 1H),6.96-6.94 (m, 1H), 6.87 (d, 1H), 6.81-6.77 (m, 1H), 3.12 (s, 3H), 2.97(s, 3H) 51

[M − H] 329 (400 MHz, CDCl₃): δ 8.25 (d, 1H), 7.33-7.30 (m, 1H),7.22-7.20 (m, 1H), 7.16-7.12 (m, 1H), 6.93-6.89 (m, 1H), 3.13 (s, 3H),2.98 (s, 3H) 52

[M − H] 322 (400 MHz, CDCl₃): δ 8.20 (d, 1H), 6.91-6.88 (m, 1H),6.81-6.75 (m, 1H), 6.72-6.65 (m, 2H), 3.12 (s, 3H), 2.97 (s, 3H) 53

[M − H] 363 (400 MHz, CDCl₃): δ 8.18 (br d, 1H), 7.26 (t, 1H), 7.01-6.97(m, 1H), 6.89- 6.84 (m, 2H), 6.71-6.67 (m, 1H), 3.15 (s, 3H), 2.95 (t,3H) 54

[M − H + 46] 425 (400 MHz, CDCl₃): δ 8.21 (d, 1H), 7.31 (t, 1H),7.03-6.98 (m, 2H), 6.89- 6.87 (m, 1H), 6.74-6.70 (m, 1H), 5.27 (d, 2H),3.30 (s, 3H), 2.96-2.91 (m, 1H) 55

[M − H] 393 (400 MHz, CDCl₃): δ 7.84 (d, 1H), 6.95 (d, 1H), 6.76-6.70(m, 1H), 6.66- 6.60 (m, 2H), 5.65-5.60 (m, 1H), 3.25-3.15 (m, 2H),3.00-2.92 (m, 1H) 2.47-2.28 (m, 2H) 56

[M − H + 46] 455 (400 MHz, CDCl₃): δ 7.84 (d, 1H), 7.03-6.99 (m, 1H),6.93 (d, 1H), 6.92- 6.90 (m, 1H), 6.75-6.71 (m, 1H), 5.65-5.61 (m, 1H),3.24-3.15 (m, 2H), 3.01-2.92 (m, 1H) 2.47-2.28 (m, 2H) 57

[M − H + 46] 446 (400 MHz, CDCl₃): δ 7.88 (d, 1H), 7.28-7.25 (m, 2H),7.19-7.17 (m, 1H), 7.09-7.05 (m, 1H), 6.96 (d, 1H), 5.66- 5.62 (m, 1H),3.23-3.13 (m, 2H), 2.99-2.90 (m, 1H) 2.47-2.29 (m, 2H) 58

[M − H] 329 (400 MHz, CDCl₃): δ 7.53-7.49 (m, 1H), 6.98-6.95 (m, 1H),6.62-6.55 (m, 1H), 6.53-6.46 (m, 2H), 5.53 (br s, 1H), 3.11-3.01 (m,1H), 2.84-2.76 (m, 1H), 2.41-2.31 (m, 1H) 2.25-2.18 (m, 1H), 2.04 (br s,1H) 59

[M − H + 46] 403 (400 MHz, CDCl₃): δ 7.81 (d, 1H), 6.97 (d, 1H),6.70-6.64 (m, 1H), 6.61- 6.55 (m, 2H), 5.70-5.66 (m, 1H), 5.41-5.14 (m,2H), 3.29 (d, 1H), 3.18- 3.09 (m, 1H), 2.92-2.83 (m, 1H), 2.51-2.42 (m,1H) 2.27-2.19 (m, 1H) 60

[M − H + 46] 403 (400 MHz, CDCl₃): δ 7.81 (d, 1H), 6.97 (d, 1H),6.70-6.64 (m, 1H), 6.61- 6.55 (m, 2H), 5.70-5.66 (m, 1H), 5.42-5.13 (m,2H), 3.30 (d, 1H), 3.18- 3.09 (m, 1H), 2.92-2.83 (m, 1H), 2.51-2.42 (m,1H) 2.27-2.19 (m, 1H) 61

[M − H + 46] 419 (400 MHz, CDCl₃): δ 7.81 (d, 1H), 6.97-6.93 (m, 2H),6.87-6.85 (m, 1H), 6.71-6.67 (m, 1H), 5.71-5.66 (m, 1H), 5.42-5.13 (m,2H), 3.30 (d, 1H), 3.18- 3.09 (m, 1H), 2.92-2.84 (m, 1H), 2.51-2.41 (m,1H) 2.28-2.19 (m, 1H) 62

[M − H + 46] 410 (400 MHz, CDCl₃): δ 7.85 (d, 1H), 7.23-7.19 (m, 2H),7.13-7.11 (m, 1H), 7.04-7.00 (m, 1H), 6.98 (d, 1H), 5.72- 5.67 (m, 1H),5.44-5.12 (m, 2H), 3.29 (d, 1H), 3.16-3.07 (m, 1H), 2.90-2.81 (m, 1H),2.52-2.42 (m, 1H), 2.29-2.20 (m, 1H) 63

[M − H + 46] 446 (400 MHz, CDCl₃): δ 7.93 (d, 1H), 7.30-7.26 (m, 1H),7.20-7.19 (m, 1H), 7.10-7.07 (m, 1H), 7.00 (d, 1H), 5.59- 5.13 (m, 3H),3.58-3.38 (m, 1H) 64

[M − H + 46] 439 (400 MHz, CDCl₃): δ 7.90 (d, 1H), 7.01 (d, 1H),6.77-6.71 (m, 1H), 6.67- 6.60 (m, 2H), 5.58-5.12 (m, 3H), 3.58-3.38 (m,3H) 65

[M − H + 46] 455 (400 MHz, CDCl₃): δ 7.90 (d, 1H), 7.03-7.00 (m, 1H),6.98 (d, 1H), 6.91- 6.90 (m, 1H), 6.76-6.72 (m, 1H), 5.58-5.12 (m, 3H),3.59-3.39 (m, 3H) 66

[M − H] 286 (400 MHz, CDCl₃): δ 7.55-7.52 (m, 1H), 6.90 (d, 1H),6.65-6.60 (m, 1H), 6.55-6.49 (m, 2H), 5.56-5.51 (m, 1H), 3.08-3.00 (m,1H), 2.80-2.71 (m, 1H), 2.68-2.64 (m, 1H) 2.60-2.50 (m, 1H), 2.17-2.08(m, 1H) 67

[M − H + 46] 368 (400 MHz, CDCl₃): δ 7.62 (d, 1H), 6.94 (d, 1H),6.72-6.67 (m, 1H), 6.61- 6.54 (m, 2H), 5.36-5.30 (m, 1H), 3.54-3.30 (m,2H), 3.13-3.10 (m, 1H) 68

[M + H] 372 (400 MHz, CDCl₃): δ 7.87 (d, 1H), 7.25-7.22 (m, 1H),7.15-7.13 (m, 1H), 7.08-6.97 (m, 2H), 5.86-5.80 (m, 1H), 3.51 (s, 3H),3.19-3.06 (m, 2H), 2.95- 2.78 (m, 1H), 2.65-2.55 (m, 1H), 2.27-2.14 (m,1H) 69

414, 416, 418 (M + H) (400 MHz, CDCl₃): δ 8.34 (d, 1H), 7.96 (m, 1H),7.08 (d, 1H), 6.99 (m, 1H), 6.86 (m, 1H), 6.70 (m, 1 H), 6.16 (t, 1H),3.35 (br s, 1H) 70

428, 430, 432 (M + H) (400 MHz, CDCl₃): δ 8.26 (d, 1H), 7.87 (m, 1H),7.07 (d, 1H), 6.98 (m, 1H), 6.86 (m, 1H), 6.70 (m, 1H), 6.22 (t, 1H),2.98 (s, 3H) 71

430, 432, 434 (M − H) (400 MHz, CDCl₃): δ 8.42 (d, 1H), 8.03 (m, 1H),7.07 (d, 1H), 7.01 (m, 1H), 6.89 (m, 1H), 6.73 (m, 1 H), 3.65 (br s, 1H)72

377, 379 (M − H) (400 MHz, CDCl₃): δ 8.47 (d, 1H), 8.23 (m, 1H), 7.12(m, 1H), 7.07 (d, 1H), 7.00 (m, 1H), 6.84 (m, 1 H), 3.74 (br s, 1H) 73

368 (M − H) (400 MHz, CDCl₃): δ 8.50 (d, 1H), 8.28 (m, 1H), 7.38 (m,1H), 7.30 (m, 1H), 7.20 (m, 1H), 7.09 (d, 1 H), 3.78 (br s, 1H) 74

462, 464, 466 (M + NH₄) (400 MHz, CDCl₃): δ 8.06 (d, 1H), 7.04-6.99 (m,2H), 6.90 (m, 1H), 6.73 (m, 1H), 6.48 (t, 1H), 5.25 (d, 2 H), 2.69 (t,1H) 75

464, 466, 468 (M − H) (400 MHz, CDCl₃): δ 8.33 (d, 1H), 7.03 (m, 1H),6.98 (d, 1H), 6.90 (m, 1H), 6.74 (m, 1H), 3.88 (br s, 1H) 76

441, 443, 445 (M + H) (400 MHz, CDCl₃): δ 8.00 (d, 1H), 7.36 (d, 1H),7.03 (m, 1H), 6.87 (m, 1H), 6.72 (m, 1H), 6.33 (t, 1H) 77

432, 434 (M + H) (400 MHz, CDCl₃): δ 8.06 (d, 1H), 7.42 (d, 1H), 7.28(m, 1H), 7.12 (m, 1H), 7.03 (m, 1H), 6.36 (t, 1H) 78

401 (M + NH₄) (400 MHz, CDCl₃): δ 7.86 (d, 1H), 7.16 (m, 1H), 7.04 (m,1H), 6.97 (d, 1H), 6.37 (t, 1H), 5.69-5.65 (m, 1H), 3.21-3.11 (m, 2H),2.92 (m, 1H), 2.51-2.41 (m, 1H), 2.32-2.23 (m, 1H) 79

(400 MHz, CDCl₃): δ 8.16 (d, 1H), 7.43 (m, 1H), 7.34-7.32 (m, 1H),7.24-7.21 (m, 1H), 7.06 (d, 1H), 2.79 (s, 3H) 80

453, 455 (M + NH₄) (400 MHz, CDCl₃): δ 8.11 (d, 1H), 7.28-7.23 (m, 1H),7.15-7.13 (m, 1H), 7.09 (d, 1H), 7.05 (m, 1H), 6.50 (t, 1H), 5.25 (d,2H), 2.69 (t, 1H) 81

435, 437 (M + NH₄) (400 MHz, CDCl₃): δ 8.05 (d, 1H), 7.59-7.56 (m, 2H),7.39-7.37 (m, 1H), 7.36-7.31 (m, 1H), 6.95 (d, 1H), 6.48 (t, 1H), 5.26(d, 2H), 2.70 (t, 1H) 82

457, 459, 461 (M + H) (400 MHz, CDCl₃): δ 8.05 (d, 1H), 7.28 (d, 1H),6.96 (m, 1H), 6.83-6.81 (m, 1H), 6.66 (m, 1H), 6.58 (m, 1H), 4.04 (t,3H) 83

488, 490, 492 (MH⁺—C₄H₈] (400 MHz, CDCl₃): δ 8.04 (d, 1H), 7.02 (m, 1H),6.99 (d, 1H), 6.90-6.88 (m, 1H), 6.73 (m, 1H), 6.62 (br t, 1H), 5.22 (brs, 1H), 4.95 (d, 2H), 1.45 (s, 9H) 84

393, 395 (M + H) (400 MHz, CDCl₃): δ 8.07 (d, 1H), 7.11 (m, 1H),7.04-6.99 (m, 2H), 6.87 (m, 1H), 6.26 (t, 1H), 5.61 (d, 2H) 85

444, 446, 448 (M + H) (400 MHz, CDCl₃): δ 8.04 (d, 1H), 7.02 (m, 1H),6.97 (d, 1H), 6.89 (m, 1H), 6.73 (m, 1H), 6.66 (t, 1H), 4.45 (br s, 2H)86

486, 488, 490 (M + H) (400 MHz, CDCl₃): δ 8.04 (d, 1H), 7.03 (m, 1H),6.99 (d, 1H), 6.90 (m, 1H), 6.74 (m, 1H), 6.66 (t, 1H), 6.11 (br s, 1H),5.05 (d, 2H), 2.00 (s, 3H) 87

379, 381 (M + H − 16) (400 MHz, CDCl₃): δ 8.07 (d, 1H), 7.02 (d, 1H),7.00 (m, 1H), 6.90-6.88 (m, 1H), 6.75-6.71 (m, 1H), 6.46 (t 1H), 5.18(d, 2H), 5.01 (d, 2H) 3.01 (t, 1H), 2.76 (t, 1H) 88

377, 379 (M + H − 16) (400 MHz, CDCl₃): δ 7.89 (d, 1H), 7.03 (m, 1H),6.97-6.94 (m, 2H), 6.80 (m, 1H), 6.67 (m, 1H), 6.20 (t, 1H), 5.57 (m,1H), 5.39 (d, 1H), 3.33 (d, 1H) 89

478, 480 (M + NH₄) (400 MHz, CDCl₃): δ 8.01 (d, 1H), 7.63-7.55 (m, 2H),7.41-7.38 (m, 1H), 7.28 (m, 1H), 6.90 (d, 1H), 6.47 (t, 1H), 5.26 (d,2H), 2.73 (t, 1H) 90

339, 341 (M + H − 16) (400 MHz, CDCl₃): δ 7.80 (d, 1H), 6.95 (d, 1H),6.93 (m, 1H), 6.84-6.82 (m, 1H), 6.66 (m, 1H), 5.68 (m, 1H), 3.64 (d,1H), 3.20 (s, 3H), 3.15-3.06 (m, 1H), 2.83 (m, 1H), 2.53-2.43 (m, 1H),2.27-2.18 (m, 1H) 91

353, 355 (M − OH) (400 MHz, CDCl₃): δ 7.74 (d, 1H), 6.95-6.92 (m, 2H),6.84-6.82 (m, 1H), 6.66 (m, 1H), 5.65-5.60 (m, 1H), 3.70 (d, 1H),3.35-3.19 (m, 2H), 3.15-3.06 (m, 1H), 2.83 (m, 1H), 2.49-2.39 (m, 1H),2.27-2.19 (m, 1H), 1.34 (t, 3H) 92

393, 395 (M + NH₄) (400 MHz, CDCl₃): δ 8.29-8.25 (m, 1H), 7.27-7.23 (m,1H), 7.22 (t, 1H), 7.10-7.06 (m, 1H), 6.93-6.91 (m, 1H), 6.76 (m, 1H),3.35 (s, 3H) 93

377 (M + NH₄) (400 MHz, CDCl₃): δ 8.29-8.25 (m, 1H), 7.29-7.25 (m, 1H),7.22 (t, 1H), 6.80 (tt, 1H), 6.69-6.63 (m, 2H), 3.35 (s, 3H) 94

384 (M + NH₄) (400 MHz, CDCl₃): δ 8.34-8.30 (m, 1H), 7.35-7.32 (m, 1H),7.29-7.25 (m, 1H), 7.21 (t, 1H), 7.21-7.18 (m, 1H), 7.11 (m, 1H), 3.36(s, 3H) 95

379 (M + NH₄) (400 MHz, CDCl₃): δ 7.98 (d, 1H), 7.17 (m, 1H), 7.05-7.03(m, 1H), 6.97 (m, 1H), 6.95 (d, 1H), 5.44-5.39 (m, 1H), 3.72 (m, 1H),3.25 (s, 3H), 3.04- 2.95 (m, 1H), 2.58-2.47 (m, 1H), 2.29-2.22 (m, 1H),2.16-2.03 (m, 1H), 1.91-1.73 (m, 2H) 96

(400 MHz, CDCl₃): δ 7.85 (d, 1H), 6.94 (d, 1H), 6.93-6.90 (m, 1H), 6.74-6.73 (m, 1H), 6.61-6.57 (m, 1H) 97

(400 MHz, CDCl₃): δ 7.68 (d, 1H), 6.99-6.94 (m, 2H), 6.85-6.84 (m, 1H),6.71-6.67 (m, 1H) 98

(M − H) 456, 458 (400 MHz, CDCl₃): δ 8.12 (d, 1H), 7.18 (d, 1H),7.14-7.11 (m, 1H), 6.97- 6.96 (m, 1H), 6.82-6.79 (m, 1H) 99

(M + NH₄) 448 (400 MHz, CDCl₃): δ 8.12 (d, 1H), 7.69-7.63 (m, 2H),7.46-7.45 (m, 1H), 7.41-7.38 (m, 1H), 7.11 (d, 1H) 100

(M + NH₄) 466 (400 MHz, CDCl₃): δ 8.17 (d, 1H), 7.39-7.36 (m, 1H),7.24-7.23 (m, 1H), 7.22 (d, 1H), 7.16-7.13 (m, 1H) 101

(400 MHz, CDCl₃): δ 10.31 (s, 1H), 7.99 (d, 1H), 7.10 (d, 1H), 7.10-7.07(m, 1H), 6.96-6.94 (m, 1H), 6.81-6.77 (m, 1H) 102

(400 MHz, CDCl₃): δ 7.77 (d, 1H), 7.16 (d, 1H), 7.00-6.97 (m, 1H), 6.83-6.82 (m, 1H), 6.70-6.67 (m, 1H), 5.43 (s, 2H) 103

(M − H) 475, 477 (400 MHz, CDCl₃): δ 8.11 (d, 1H), 7.06-7.03 (m, 1H),6.95 (d, 1H), 6.92- 6.91 (m, 1H), 6.77-6.74 (m, 1H), 5.88 (m, 1H), 3.38(d, 1H), 1.81 (d, 3H) 104

(M + H) 474.8/ 476.7 (400 MHz, CDCl₃): δ 7.97-7.94 (m, 1H), 7.10-7.07(m, 1H), 7.01 (d, 1H), 6.80-6.77 (m, 1H), 2.71 (s, 3H) 105

(M + H) 424, 426 (400 MHz, CDCl₃): δ 8.35 (d, 1H), 7.84 (brd s, 1H),7.26 (d, 1H), 7.15- 7.12 (m, 1H), 7.04-7.03 (m, 1H), 6.89-6.86 (m, 1H)106

(M + H) 492, 494 (400 MHz, CDCl₃): δ 7.95-7.94 (m, 1H), 7.09-7.06 (m,1H), 7.01 (d, 1H), 6.96-6.95 (m, 1H), 6.80-6.77 (m, 1H), 2.70 (s, 3H)107

(M + H) 476, 478 (400 MHz, CDCl₃): δ 8.39 (s, 1H), 8.07 (d, 1H), 7.07(d, 1H), 7.07-7.04 (m, 1H), 6.94-6.93 (m, 1H), 6.79-6.76 (m, 1H) 108

(M + H) 506, 508 (400 MHz, CDCl₃): δ 8.07 (d, 1H), 7.06-7.03 (m, 1H),6.98 (d, 1H), 6.93- 6.92 (m, 1H), 6.78-6.74 (m, 1H), 4.32 (s, 2H), 3.72(t, 2H), 2.97 (t, 2H) 109

(M + H) 476, 478 (400 MHz, CDCl₃): δ 8.06 (d, 1H), 7.05-7.02 (m, 1H),6.97 (d, 1H), 6.91- 6.90 (m, 1H), 6.76-6.72 (m, 1H), 4.25 (s, 2H), 2.57(s, 3H) 110

(M + H) 552, 554 (400 MHz, CDCl₃): δ 8.04 (d, 1H), 7.39-7.31 (m, 4H),7.28-7.23 (m, 1H), 7.05-7.02 (m, 1H), 6.94 (d, 1H), 6.91- 6.89 (m, 1H),6.75-6.72 (m, 1H), 4.30 (s, 2H), 3.96 (s, 2H). 111

(M + NH₄) 431, 433 (400 MHz, CDCl₃): δ 8.08 (d, 1H), 7.23 (d, 1H),7.14-7.11 (m, 1H), 6.98- 6.96 (m, 1H), 6.83-6.79 (m, 1H) 112

(400 MHz, CDCl₃): δ 10.43 (s, 1H), 7.96 (d, 1H), 7.14 (d, 1H), 7.09-7.07(m, 1H), 6.95-6.94 (m, 1H), 6.80-6.77 (m, 1H) 113

(M + H) 432, 434 (400 MHz, CDCl₃): δ 8.02 (d, 1H), 7.05-7.02 (m, 1H),7.01 (d, 1H), 6.91- 6.90 (m, 1H), 6.76-6.72 (m, 1H), 4.22 (s, 2H), 2.56(s, 3H) 114

(M + H) 464, 466 (400 MHz, CDCl₃): δ 8.02 (d, 1H), 7.06-7.03 (m, 1H),7.01 (d, 1H), 6.92 (m, 1H), 6.77-6.73 (m, 1H), 4.64 (t, 1H), 4.52 (t,1H), 4.34 (s, 2H), 3.11- 3.09 (m, 1H), 3.04-3.02 (m, 1H) 115

(M + H) 376, 378 (400 MHz, CDCl₃): δ 8.50-8.49 (m, 1H), 8.36-8.35 (m,1H), 7.89 (d, 1H), 7.43 (t, 1H), 6.93 (d, 1H), 5.62-5.58 (m, 1H),3.62-3.40 (m, 3H), 3.22 (s, 3H) 116

431 (M − H) (400 MHz, CDCl₃): δ 8.30 (d, 1 H), 7.93 (m, 1 H), 7.08-7.02(m, 2 H), 6.93-6.91 (m, 1 H), 6.78-6.74 (m, 1 H) 117

404 (M − H) 118

422 (M − H) 119

379 (M − H) (400 MHz, CDCl₃): δ 8.35 (d, 1 H), 8.13 (m, 1 H), 7.16-7.13(m, 1 H), 7.11 (d, 1 H), 7.03-7.01 (m, 1 H), 6.88-6.85 (m, 1 H) 120

369 (M − H) 121

411 (M − H) (400 MHz, CDCl₃): δ 8.58 (d, 1 H), 8.10-8.07 (m, 1 H), 7.14(d, 1 H), 7.03-7.00 (m, 1 H), 6.91-6.90 (m, 1 H), 6.77-6.73 (m, 1 H),3.94 (s, 3 H) 122

383 (M − H) 123

124

(400 MHz, CDCl₃): δ 8.35-8.34 (m, 1 H), 8.09-8.05 (m, 1 H), 7.17-6.90(m, 4 H), 6.82-6.78 (m, 1 H) 125

396 (M − H) (400 MHz, CDCl₃): δ 8.95 (d, 1 H), 8.07-8.04 (m, 1 H), 7.21(br s, 1 H), 7.15-7.12 (m, 1 H), 7.05 (d, 1 H), 7.02-7.01 (m, 1 H),6.86-6.83 (m, 1 H), 6.01 (br s, 1 H) 126

457 (M + HCO₂ ⁻) (400 MHz, CDCl₃): δ 7.96 (d, 1 H), 7.84 (m, 1 H),7.02-6.98 (m, 2 H), 6.88-6.87 (m, 1 H), 6.73-6.69 (m, 1 H), 3.74-3.60(m, 2 H), 2.91-2.87 (m, 2 H), 1.97-1.90 (m, 2 H), 1.40-1.37 (m, 1 H) 127

397 (M − H) (400 MHz, CDCl₃): δ 8.02 (d, 1 H), 7.86 (m, 1 H), 7.02-6.99(m, 2 H), 6.89-6.87 (m, 1 H), 6.74-6.70 (m, 1 H), 3.98-3.93 (m, 2 H),3.06 (t, 2 H), 1.50-1.47 (m, 1 H) 128

387 (M − H) 129

421 (M − H) (400 MHz, CDCl₃): δ 8.39-8.38 (m, 1 H), 7.98-7.95 (m, 1 H),7.15 (d, 1 H), 7.08-7.05 (m, 1 H), 6.95-6.94 (m, 1 H), 6.80-6.77 (m, 1H) 130

389 (M − H) (400 MHz, CDCl₃): δ 8.18 (d, 1 H), 7.83-7.80 (m, 1 H), 7.07(d, 1 H), 6.97-6.94 (m, 1 H), 6.82-6.81 (m, 1 H), 6.70-6.64 (m, 2 H),6.54-6.50 (m, 1H), 6.13-6.11 (m, 1H) 131

413 (M − H) (400 MHz, CDCl₃): δ 8.26 (d, 1 H), 7.89-7.87 (m, 1 H), 7.07(d, 1 H), 7.04-7.00 (m, 1 H), 6.90-6.89 (m, 1 H), 6.75-6.72 (m, 1 H),6.21 (t, 1 H) 132

404 (M − H) (400 MHz, CDCl₃): δ 8.30 (d, 1 H), 7.95-7.93 (m, 1 H),7.27-7.25 (m, 1 H), 7.15-7.13 (m, 2 H), 7.06-7.03 (m, 1 H), 6.24 (t, 1H) 133

360 (M − H) (400 MHz, CDCl₃): δ 8.34 (d, 1 H), 8.15-8.12 (m, 1 H),7.40-7.37 (m, 1 H), 7.31-7.29 (m, 1 H), 7.22-7.19 (m, 1 H), 7.10 (d, 1H), 6.26 (t, 1 H) 134

351 (M − H) (400 MHz, CDCl₃): δ 8.34 (d, 1 H), 8.15-8.12 (m, 1 H),7.40-7.37 (m, 1 H), 7.31-7.29 (m, 1 H), 7.22-7.19 (m, 1 H), 7.10 (d, 1H), 6.26 (t, 1 H) 135

349 (M − H) (400 MHz, CDCl₃): δ 7.86 (d, 1 H), 7.80-7.77 (m, 1 H), 7.01(d, 1 H), 6.98-6.95 (m, 1 H), 6.84-6.83 (m, 1 H), 6.69-6.65 (m, 1 H),6.19 (t, 1 H), 2.39 (s, 3 H) 136

340 (M − H) (400 MHz, CDCl₃): δ 7.93 (d, 1 H), 7.85-7.82 (m, 1 H),7.23-7.20 (m, 1H), 7.11-7.09 (m, 1 H), 7.04 (d, 1 H), 7.03-6.98 (m, 1H), 6.21 (t, 1 H), 2.39 (s, 3 H) 137

395 (M − H) (400 MHz, CDCl₃): δ 8.25 (d, 1 H), 7.89-7.86 (m, 1 H), 7.09(d, 1 H), 7.00-6.97 (m, 1 H), 6.87-6.86 (m, 1 H), 6.72-6.69 (m, 1 H),5.17 (d, 2 H) 138

377 (M − H) 139

370 (M + HCO₂ ⁻) (400 MHz, CDCl₃): δ 8.29-8.28 (m, 1 H), 8.09-8.06 (m, 1H), 7.10-7.06 (m, 2 H), 6.97-6.96 (m, 1 H), 6.83-6.79 (m, 1 H), 3.10 (s,3 H) 140

445 (M − H) (400 MHz, CDCl₃): δ 8.07 (d, 1 H), 7.03-7.00 (m, 1 H), 6.94(d, 1 H), 6.87-6.86 (m, 1 H), 6.72-6.68 (m, 1 H), 2.52 (s, 3 H) 141

438 (M + H) 142

412 (M − H) (400 MHz, CDCl₃): δ 8.00 (d, 1 H), 7.26-7.23 (m, 1 H),7.12-7.11 (m, 1 H), 7.03-6.99 (m, 1 H), 6.97 (d, 1 H), 4.62 (d, 2 H),2.49 (s, 3 H), 1.96-1.91 (m, 1 H) 143

401 (M − H) (400 MHz, CDCl₃): δ 8.03 (d, 1 H), 7.03-7.00 (m, 1 H), 6.90(d, 1 H), 6.88-6.86 (m, 1 H), 6.72-6.69 (m, 1 H), 2.47 (s, 3 H) 144

392 (M − H) (400 MHz, CDCl₃): δ 8.05 (m, 1 H), 7.29-7.26 (m, 1 H), 7.14(s, 1 H), 7.05-7.02 (m, 1 H), 6.94-6.91 (m, 1 H), 2.46 (s, 3 H) 145

386 (M − H) (400 MHz, CDCl₃): δ 10.62 (s, 1 H), 7.97 (d, 1 H), 7.30-7.27(m, 1 H), 7.16-7.15 (m, 1 H), 7.10 (d, 1 H), 7.07-7.03 (m, 1 H), 2.40(s, 3 H) 146

434 (M + HCO₂ ⁻) (400 MHz, CDCl₃): δ 8.01 (d, 1 H), 7.26-7.23 (m, 1 H),7.12-7.11 (m, 1 H), 7.03-6.99 (m, 2 H), 4.99 (d, 2 H), 2.50 (s, 3 H) 147

442 (M − H) (400 MHz, CDCl₃): δ 8.02 (s, 1 H), 7.99 (d, 1 H), 7.28-7.25(m, 1 H), 7.16-7.15 (m, 1 H), 7.07-7.05 (m, 1 H), 6.98 (d, 1 H), 6.03(d, 1 H), 3.85 (s, 3 H), 2.34 (s, 3 H) 148

458 (M + H) (400 MHz, CDCl₃): δ 8.02 (s, 1 H), 7.99 (d, 1 H), 7.28-7.25(m, 1 H), 7.16-7.15 (m, 1 H), 7.07-7.03 (m, 1 H), 6.98 (d, 1 H), 6.02(d, 1 H), 4.31 (q, 2 H), 2.34 (s, 3 H), 1.36 (t, 3 H) 149

519 (M + H) (400 MHz, CDCl₃): δ 8.01 (d, 1 H), 7.79 (d, 1 H), 7.39-7.29(m, 5 H), 7.27-7.24 (m, 1 H), 7.14-7.13 (m, 1 H), 7.06-7.03 (m, 1 H),6.97 (d, 1 H), 6.06-6.02 (m, 2 H), 4.60 (d, 2 H), 2.33 (s, 3 H) 150

454 (M + H) (400 MHz, CDCl₃): δ 8.47 (d, 1 H), 8.05 (d, 1 H), 7.98 (d, 1H), 7.31-7.27 (m, 1 H), 7.19-7.18 (m, 1 H), 7.10- 7.07 (m, 1 H), 7.01(d, 1 H), 6.72 (d, 1 H), 2.42 (s, 3 H) 151

468 (M + H) (400 MHz, CDCl3): δ 8.03 (d, 1 H), 7.85 (d, 1 H), 7.30-7.27(m, 1 H), 7.18-7.17 (m, 1 H), 7.09-7.05 (m, 1 H), 6.99 (d, 1 H), 6.62(d, 1 H), 2.63 (s, 3 H), 2.40 (s, 3 H) 152

349 (M − H) (400 MHz, CDCl3): δ 8.32 (d, 1 H), 7.38-7.35 (m, 1 H),7.26-7.25 (m, 1 H), 7.18-7.14 (m, 1 H), 6.97 (d, 1 H), 3.30 (s, 3 H) 153

396 (M − H) (400 MHz, CDCl3): δ 8.01 (d, 1 H), 7.22 (d, 1 H), 7.09-7.05(m, 1 H), 6.94-6.92 (m, 1 H), 6.8-6.77 (m, 1 H) 154

437 (M − H) (400 MHz, CDCl3): δ 7.44-7.40 (m, 1 H), 7.33 (m, 1 H), 7.15(d, 1 H), 6.91-6.88 (m, 1 H), 6.74-6.73 (m, 1 H), 6.62-6.58 (m, 1 H),3.95-3.91 (m, 2 H), 3.44-3.40 (m, 2 H), 1.71-1.69 (m, 1 H) 155

428 (M + HCO₂ ⁻) (400 MHz, CDCl₃): δ 7.85 (d, 1 H), 7.26-7.24 (m, 1 H),7.17-7.15 (m, 1 H), 7.06-7.03 (m, 1 H), 6.97 (d, 1 H), 6.37 (t, 3 H),5.68-5.65 (m, 1 H), 3.20-3.11 (m, 2 H), 2.94-2.87 (m, 1 H), 2.51-2.41(m, 1 H), 2.31-2.24 (m, 1 H) 156

410 (M + HCO₂ ⁻) (400 MHz, CDCl₃): δ 7.80 (d, 1 H), 7.56-7.54 (m, 2 H),7.39-7.30 (m, 2 H), 6.88-6.84 (m, 1 H), 6.38 (t, 1 H), 5.68-5.66 (m, 1H), 3.22-3.13 (m, 2 H), 2.98-2.90 (m, 1 H), 2.50-2.41 (m, 1 H),2.32-2.22 (m, 1 H) 157

437 (M + HCO₂ ⁻) 158

473 (M + HCO₂ ⁻) (400 MHz, CDCl₃): δ 7.90 (d, 1 H), 7.06-7.03 (m, 1 H),6.99 (d, 1 H), 6.94-6.93 (m, 1 H), 6.78-6.75 (m, 1 H), 6.43 (t, 1 H),5.52-5.48 (m, 1 H), 3.64-3.43 (m, 2 H), 3.29 (s, 1 H) 159

421 (M + HCO₂ ⁻) (400 MHz, CDCl₃): δ 7.89 (d, 1 H), 7.01 (d, 1 H),6.74-6.68 (m, 1 H), 6.62-6.58 (m, 2 H), 5.61-5.57 (m, 1H), 3.54-3.40 (m,3 H), 3.22 (s, 3 H) 160

393 (M + H) (400 MHz, CDCl₃): δ 7.90-7.87 (m, 1 H), 7.01-6.97 (m, 2 H),6.88-6.87 (m, 1 H), 6.73-6.69 (m, 1 H), 5.61-5.57 (m, 1 H), 3.57-3.37(m, 3 H), 3.22 (s, 3 H) 161

421 (M + HCO₂ ⁻) (400 MHz, CDCl₃): δ 7.89 (d, 1 H), 7.01 (d, 1 H),6.74-6.68 (m, 1 H), 6.62-6.58 (m, 2 H), 5.61-5.57 (m, 1H), 3.54-3.40 (m,3 H), 3.22 (s, 3 H) 162

410 (M + HCO₂ ⁻) 163

428 (M + HCO₂ ⁻) (400 MHz, CDCl₃): δ 7.93 (d, 1 H), 7.27-7.24 (m, 1 H),7.15-7.14 (m, 1 H), 7.07-7.03 (m, 1 H), 7.00 (d, 1 H), 5.63-5.58 (m, 1H), 3.56-3.35 (m, 3 H), 3.24 (s, 3 H) 164

383 (M + H) (400 MHz, CDCl₃): δ 7.93-7.91 (m, 1 H), 7.25-7.22 (m, 1 H),7.14-7.13 (m, 1 H), 7.06-7.02 (m, 1 H), 6.96 (d, 1 H), 4.97-4.93 (m, 1H), 3.55-3.37 (m, 2 H), 3.32 (s, 3 H) 165

383 (M + H) (400 MHz, CDCl₃): δ 7.93-7.91 (m, 1 H), 7.25-7.22 (m, 1 H),7.14-7.13 (m, 1 H), 7.06-7.02 (m, 1 H), 6.96 (d, 1 H), 4.97-4.93 (m, 1H), 3.55-3.37 (m, 2 H), 3.32 (s, 3 H) 166

[M − H + 46]: 442 (400 MHz, CDCl₃) δ 7.87 (d, 1H), 7.38 (br s, 1H), 7.16(br d, 1H), 6.88 (d, 1H), 5.58-5.12 (m, 3H), 3.59-3.44 (m, 3H) 167

[M − H + 46]: 462 (400 MHz, CDCl₃) δ 7.92 (d, 1H), 7.36-7.33 (m, 1H),7.32-7.27 (m, 1H), 6.97 (d, 1H), 5.58-5.12 (m, 3H), 3.62- 3.38 (m, 3H)168

[M − H + 46]: 429 (400 MHz, CDCl₃) δ 8.01 (d, 1H), 7.09 (d, 1H),6.77-6.67 (m, 1H), 6.64- 6.61 (m, 2H), 6.47 (t, 1H), 5.21 (d, 2H), 2.70(t, 1H) 169

(400 MHz, CDCl₃): δ 8.30 (d, 1H), 7.89 (d, 1H), 7.54 (d, 1H), 7.26 (s,1H), 6.99 (m, 2H) 170

(400 MHz, CDCl₃): δ 8.30 (s, 1H), 7.88 (d, 1H), 7.21-7.31 (m, 3H), 6.90(d, 1H) 171

(400 MHz, d₆-DMSO): δ 8.40 (d, 1H), 8.02 (d, 1H), 7.45 (d, 1H), 7.35 (t,1H), 7.20 (d, 1H), 6.95 (d, 1H), 2.13 (s, 3H) 172

(400 MHz, CDCl₃): δ 8.30 (d, 1H), 7.93 (d, 1H), 7.31 (d, 1H), 7.04 (m,3H) 173

(400 MHz, CDCl₃): δ 8.26 (d, 1H), 7.81 (d, 1H), 6.95 (s, 1H), 6.91 (d,1H), 6.74 (s, 2H), 2.35 (s, 6H) 174

(400 MHz, CDCl₃): δ 8.29 (d, 1H), 7.84 (d, 1H), 7.22-7.27 (m, 2H), 7.06(s, 1H), 6.78 (d, 1H), 2.16 (s, 3H) 175

(400 MHz, d₆-DMSO): δ 8.40 (s, 1H), 8.03 (d, 1H), 7.58 (m, 2H), 7.23 (m,1H), 7.11 (d, 1H) 176

(400 MHz, d₆-DMSO): δ 8.30 (s, 1H), 7.88 (d, 1H), 7.27-7.32 (m, 1H),7.08- 7.14 (m, 2H), 6.85 (d, 1H) 177

(400 MHz, CDCl₃): δ 8.29 (d, 1 H), 7.87 (m, 1 H), 7.06 (d, 1H), 7.02 (d,1H), 6.94 (d, 1H), 6.77 (m, 1H), 4.75 (d, 2H), 1.83 (t, 1H) 178

391 (M − H) (400 MHz, CDCl₃): δ 8.18 (d, 1H), 7.38-7.35 (m, 1H),7.29-7.27 (m, 1H), 7.21-7.18 (m, 1H), 7.07-7.02 (m, 1H), 6.90 (d, 1H),6.56-6.47 (m, 1H), 6.22 (t, 1H), 2.08-2.06 (m. 1H) 179

(400 MHz, CDCl₃): δ 8.13 (d, 1H), 7.08-7.05 (m ,1H), 6.95-6.93 (m, 2H),6.79-6.76 (m, 1H) 180

359, 361 (M + H) (400 MHz, CDCl₃): δ 8.33 (d, 1H), 7.09 (m, 1H),6.97-6.95 (m, 1H), 6.94 (d, 1H), 6.81 (m, 1H), 3.32 (s, 3H), 2.94 (br s,1H) 181

359, 361 (M + H) (400 MHz, CDCl₃): δ 8.04 (d, 1H), 7.23 (m, 1H),7.12-7.10 (m, 1H), 7.03-6.99 (m, 1H), 6.07 (d, 1H), 5.54- 5.49 (m, 1H),3.68 (m, 1H), 3.26 (s, 3H), 3.20 (m, 1H), 2.97-2.86 (m, 1H), 2.63-2.45(m, 1H), 2.35-2.25 (m, 1H) 182

(M + H) 410/412 183

478 mCi 184

[M + H] 394 (400 MHz, CDCl₃): δ 8.47 (d, 1H), 8.35 (d, 1H), 7.82 (d,1H), 7.45 (t, 1H), 6.88 (d, 1H), 5.64-5.59 (m, 1H), 3.30-3.15 (m, 2H),3.02-2.93 (m, 1H) 2.46-2.26 (m, 2H) 185

[M − H] 436 (400 MHz, CDCl₃): δ 7.95 (d, 1H), 7.35-7.31 (m, 1H),7.26-7.23 (m, 1H), 7.15-7.11 (m, 1H), 6.99 (d, 1H), 5.46- 5.39 (m, 1H),3.63-3.41 (m, 2H), 3.36 (d, 1H) 186

[M + H] 430 (400 MHz, CDCl₃): δ 8.55 (d, 1H), 8.40 (d, 1H), 7.91 (d,1H), 7.52 (t, 1H), 6.94 (d, 1H), 5.46-5.40 (m, 1H), 3.85 (d, 1H),3.66-3.47 (m, 2H) 187

[M + H] 394 (400 MHz, CDCl₃): δ 8.51 (d, 1H), 8.37 (d, 1H), 7.90 (d,1H), 7.48 (t, 1H), 6.93 (d, 1H), 5.61-5.11 (m, 3H), 3.94 (d, 1H),3.62-3.42 (m, 2H) 188

[M − H + 46] 457 (400 MHz, CDCl₃): δ 7.89 (d, 1H), 6.94 (d, 1H),6.82-6.71 (m, 2H), 5.59- 5.11 (m, 3H), 3.59-3.38 (m, 3H) 189

[M + H] 365 (400 MHz, CDCl₃): δ 7.88-7.80 (m, 1H), 6.97 (d, 1H),6.72-6.65 (m, 1H), 6.63-6.55 (m, 2H), 5.83-5.76 (m, 1H), 3.57 (s, 1H),3.51 (s, 3H), 3.18-3.07 (m, 1H), 2.93-2.79 (m, 1H), 2.60-2.47 (m, 1H),2.23-2.11 (m, 1H) 190

[M + H] 340 (400 MHz, CDCl₃): δ 7.89-7.81 (m, 1H), 6.99-6.94 (m, 1H),6.66-6.60 (m, 1H), 6.57-6.51 (m, 2H), 5.66-5.59 (m, 1H), 3.28 (s, 3H),3.24 (s, 1H), 3.15- 3.01 (m, 1H), 2.87-2.71 (m, 1H), 2.55-2.41 (m, 1H),2.27-2.13 (m, 1H) 191

[M + H] 401 (400 MHz, CDCl₃): δ 7.98-7.91 (m, 1H), 7.04-7.01 (m, 1H),6.80-6.73 (m, 1H), 6.69-6.61 (m, 2H), 5.73-5.63 (m, 1H), 3.60 (s, 1H),3.58-3.40 (m, 5H) 192

[M + H] 376 (400 MHz, CDCl₃): δ 7.96-7.98 (m, 1H), 7.03-6.98 (m, 1H),6.73-6.66 (m, 1H), 6.63-6.55 (m, 2H), 5.62-5.56 (m, 1H), 5.47-5.41 (m,1H), 3.57-3.30 (m, 2H), 3.28 (s, 3H), 3.24-2.88 (m, 1H) 193

(400 MHz, CDCl₃) δ 8.18 (d, 1H), 7.41-7.39 (m, 1H), 7.30-7.26 (m, 1H),7.13-7.05 (m, 2H), 6.91 (t, 1H), 3.67 (t, 2H) 194

(M + HCOOH—H): 517, 519 (400 MHz, CDCl₃) δ 7.86 (d, 1H), 7.37-7.35 (m,1H), 7.25-7.21 (m, 1H), 7.08-7.04 (m, 1H), 6.86 (d, 1H), 6.41 (t, 1H),5.51-5.47 (m, 1H), 3.63-3.47 (m, 2H), 3.25 (d, 1H) 195

(400 MHz, CDCl₃) δ 8.16 (d, 1H), 7.32 (q, 1H), 7.13 (d, 1H), 7.06-7.02(m, 1H), 6.93-6.91 (m, 1H), 6.90 (t, 1H), 3.67 (t, 2H) 196

(M + HCOOH—H) 457 (400 MHz, CDCl₃) δ 7.86 (d, 1H), 7.30-7.24 (m, 1H),7.02-6.97 (m, 1H), 6.89-6.86 (m, 2H), 6.41 (t, 1H), 5.51- 5.47 (m, 1H),3.63-3.47 (m, 2H), 3.27 (d, 1H) 197

(400 MHz, CDCl₃) δ 8.14 (d, 1H), 7.15-7.05 (m, 2H), 6.99-6.91 (m, 2H),6.92 (t, 1H), 3.67 (t, 2H), 2.33 (m, 3H) 198

(M + HCOOH—H) 453 (400 MHz, CDCl₃) δ 7.82 (d, 1H), 7.09 (t, 1H),6.93-6.88 (m, 2H), 6.82 (d, 1H), 6.40 (t, 1H), 5.48 (m, 1H), 3.63-3.48(m, 2H), 3.25 (d, 1H), 2.31 (m, 3H) 199

(400 MHz, CDCl₃) δ 8.20 (d, 1H), 7.47-7.38 (m, 3H), 7.11 (d, 1H), 6.92(t, 1H), 3.68 (t, 2H) 200

(M + HCOOH—H) 464 (400 MHz, CDCl₃) δ 7.89 (d, 1H), 7.41-7.32 (m, 3H),6.85 (d, 1H), 6.43 (t, 1H), 5.57-5.48 (m, 1H), 3.59-3.49 (m, 2H), 3.29(d, 1H) 201

(M + HCOOH—H) 401 (400 MHz, CDCl₃) δ 8.31 (s, 1H), 8.28 (s, 1H), 7.94(d, 1H), 7.33 (d, 1H), 5.61-5.58 (m, 1H), 3.57 (d, 1H), 3.51-3.28 (m,2H), 3.24 (s, 3H), 2.44 (s, 3H) 202

(M + HCOOH—H) 428 (400 MHz, CDCl₃) δ 7.95 (d, 1H), 7.28-7.26 (m, 1H),7.18 (brd s, 1H), 7.08-7.03 (m, 2H), 6.64-6.47 (m, 1H), 6.34 (t, 1H),3.23-3.14 (m, 1H), 3.04- 2.95 (m, 1H), 2.57-2.42 (m, 2H) 203

(400 MHz, CDCl₃) δ 7.78-7.75 (m, 1H), 7.05-7.02 (m, 1H), 7.00-6.98 (m,1H), 6.93-6.92 (m, 1H), 6.78-6.75 (m, 1H) 204

350, 352, 354 (M − H) (400 MHz, CDCl₃): δ 7.71 (d, 1H), 7.38 (t, 1H),7.31-7.26 (m, 1H), 7.14 (t, 1H), 7.03-7.00 (m, 1H), 6.91 (d, 1H) 205

(400 MHz, CDCl₃): δ 7.79 (d, 1H), 7.18 (d, 1H), 7.09 (s, 1H), 6.98 (m,2H), 5.65 (m, 1H), 3.69 (d, 1H), 3.29 (m, 2H), 3.08 (m, 1H), 2.83 (m,1H), 2.45 (m, 1H), 2.24 (m, 1H), 1.36 (t, 3H) 206

442 (M + HCO2) (400 MHz, CDCl₃): δ 7.86 (m, 1 H), 7.27-7.24 (m, 1 H),7.16-7.14 (m, 1 H), 7.07-7.04 (m, 1 H), 6.99 (d, 1 H), 5.55-5.51 (m, 1H), 3.61-3.27 (m, 5 H), 1.35 (t, 3 H) 207

419 (M + H) (400 MHz, CDCl₃): δ 7.94 (d, 1H), 7.32-7.28 (m, 1H),7.22-7.19 (m, 1H), 7.12-7.07 (m, 1H), 6.94 (d, 1H), 6.83 (t, 1H), 4.91(d, 1H), 3.60-3.40 (m, 2H), 1.91 (br s, 2H) 208

(400 MHz, CDCl₃): δ 7.73 (d, 1H), 7.18 (d, 1H), 7.08 (s, 1H), 6.95 (m,2H), 5.62 (m, 1H), 4.02 (m, 1H), 3.77 (s, 1H), 3.07 (m, 1H), 2.81 (m,1H), 2.61 (m, 2H), 2.45 (m 1H), 2.26 (m, 3H), 2.06 (m, 2H) 209

468 (M + HCO2) (400 MHz, CDCl₃): δ 7.81 (d, 1 H), 7.27-7.24 (m, 1 H),7.15-7.14 (m, 1 H), 7.06-7.03 (m, 1 H), 6.96 (d, 1 H), 5.50-5.45 (m, 1H), 3.70 (d, 1 H), 3.55-3.34 (m, 2 H), 2.67-2.50 (m, 2 H), 2.29-2.17 (m,2 H), 2.08-2.01 (m, 2 H) 210

398 (M + H) (400 MHz, CDCl₃): δ 7.95 (d, 1H), 7.24 (d, 1H), 7.13 (br s,1H), 7.05- 7.01 (m, 1H), 6.99 (d, 1H), 5.31 (d, 1H), 3.78 (s, 3H),3.53-3.32 (m, 2H), 3.19 (s, 3H) 211

354 (M − H) (400 MHz, CDCl₃): δ 7.75 (d, 1H), 6.94 (d, 1H), 6.65-6.60(m, 1H), 6.54- 6.52 (m, 2H), 5.77-5.71 (m, 1H), 5.02-4.95 (m, 1H),3.23-3.18 (m, 1H), 3.12-3.04 (m, 1H), 2.84-2.70 (m, 1H), 2.65 (d, 3H),2.57-2.47 (m, 1H), 2.19- 2.11 (m, 1H) 212

(400 MHz, CDCl₃): δ 7.77 (d, 1H), 7.05-7.26 (m, 4H), 6.75 (d, 1H), 5.62(m, 1H), 3.17-3.30 (m, 2H), 2.98-3.07 (m, 1H), 2.40-2.47 (m, 1H),2.28-2.37 (m, 1H) 213

(400 MHz, CDCl₃): δ 7.22-7.25 (m, 1H), 7.08 and 7.12 (m 1H), 6.98-7.04(m 1H), 6.80 (s, 1H), 5.58 and 5.78 (m 1H), 3.69 (d, 1H), 3.20 and 3.23(s, 3H), 3.08-3.47 (m, 2H), 2.68 (s, 3H) 214

(400 MHz, d₆-DMSO): 7.87 (d, 1H), 7.51-7.64 (m, 2H), 7.11-7.16 (m, 1H),6.96 (d, 1H), 5.51 (m, 1H), 5.30 (d, 1H), 3.04-3.31 (m, 1H), 2.87-2.95(m 1H), 2.11-2.30 (m, 1H), 1.99-2.09 (m, 1H) 215

(400 MHz, CDCl₃): 7.92 (d, 1H), 7.27 (m, 2H), 7.08 (d, 1H), 6.99 (d,1H), 5.63 (dd, 1H), 4.92 (d, 1H), 4.65 (d, 1H), 3.34-3.49 (m, 2H), 3.21(s, 1H) 216

(400 MHz, CDCl3): 7.83 (d. 1H), 7.26-7.38 (m, 3H), 6.81 (d, 1H), 5.64(dd, 1H), 3.16-3.25 (m, 2H), 3.00-3.04 (m 1H), 2.34-2.42 (m, 2H) 217

418 (M + HCO₂ ⁻) (400 MHz, CDCl₃): 7.88 (d, 1H), 7.17 (d, 1H), 7.09 (s,1H), 7.06 (m, 2H), 5.07 (d, 1H), 3.20 (m, 5H), 2.60 (d, 1H), 1.18-1.32(m, 2H), 0.68-0.87 (m, 2H) 218

(400 MHz, CDCl3): 7.80 (d, 1H), 7.18-7.23 (m, 2H), 6.97-7.01 (m, 1H),6.80 (d, 1H), 5.63 (m 1H), 3.16-3.29 (m, 2H), 2.96-3.05 (m 1H),2.29-2.46 (m, 2H) 219

(400 MHz, d6-DMSO): δ 7.85 (d, 1H), 7.67 (m, 1H), 7.46 (d, 1H), 6.85 (d,1H), 5.38 (dd, 1H), 3.40-3.49 (m, 2H), 3.40 (s, 3H) 220

221

445, 447 (M − H) (400 MHz, CDCl₃): δ 7.97 (d, 1 H), 7.28-7.22 (m, 2 H),7.02 (d, 1 H), 6.87 (d, 1 H), 5.43-5.39 (m, 1 H), 3.64-3.47 (m, 2 H),3.26 (d, 1 H) 222

392 (M + HCO₂ ⁻) (400 MHz, CDCl₃): δ 7.84 (d, 1 H), 7.19-7.17 (m, 1 H),7.08 (s, 1 H), 7.00-6.97 (m, 2 H), 5.71-5.68 (m, 1 H), 3.64 (d, 1 H),3.21 (s, 3 H), 3.12- 3.04 (m, 1 H), 2.84-2.76 (m, 1 H), 2.52-2.43 (m, 1H), 2.27-2.19 (m, 1 H) 223

428 (M + HCO₂ ⁻) (400 MHz, CDCl₃): δ 7.93 (d, 1 H), 7.27-7.24 (m, 1 H),7.15-7.14 (m, 1 H), 7.07-7.03 (m, 1 H), 7.00 (d, 1 H), 5.63-5.58 (m, 1H), 3.56-3.35 (m, 3 H), 3.24 (s, 3 H) 224

399 (M + H) (400 MHz, CDCl₃): δ 7.87 (d, 1 H), 7.40 (s, 1 H), 7.36 (s, 1H), 7.08 (d, 1 H), 5.42-5.38 (m, 1 H), 3.94 (s, 3 H), 3.59-3.52 (m, 2H), 3.21 (d, 1 H) 225

401 (M − H) (400 MHz, CDCl₃): δ 8.82 (d, 1 H), 8.70 (d, 1 H), 7.95 (d, 1H), 7.71-7.69 (m, 1 H), 6.94 (d, 1 H), 5.64-5.59 (m, 1 H), 5.46-5.31 (m,1 H), 3.36-3.27 (m, 2 H), 3.19 (d, 1 H) 226

428 (M + HCO₂ ⁻) (400 MHz, CDCl₃): δ 7.93 (d, 1 H), 7.27-7.24 (m, 1 H),7.15-7.14 (m, 1 H), 7.07-7.03 (m, 1 H), 7.00 (d, 1 H), 5.63-5.58 (m, 1H), 3.56-3.35 (m, 3 H), 3.24 (s, 3 H) 227

428 (M + HCO₂ ⁻) (400 MHz, CDCl₃): δ 7.93 (d, 1 H), 7.27-7.24 (m, 1 H),7.15-7.14 (m, 1 H), 7.07-7.03 (m, 1 H), 7.00 (d, 1 H), 5.63-5.58 (m, 1H), 3.56-3.35 (m, 3 H), 3.24 (s, 3 H) 228

436 (M − H) (400 MHz, CDCl₃): δ 7.90 (d, 1 H), 7.42-7.30 (m, 3 H), 6.86(d, 1 H), 5.42 (dd, 1 H), 3.58-3.47 (m, 2 H), 3.32 (d, 1 H) 229

432 (M + HCO₂ ⁻) (400 MHz, CDCl₃): δ 7.92 (d, 1 H), 7.26-7.24 (m, 1 H),7.15 (s, 1 H), 7.06-7.03 (m, 1 H), 7.01 (d, 1 H), 3.56-3.35 (m, 3 H) 230

411 (M − H) (400 MHz, CDCl₃): δ 7.85 (d, 1 H), 7.19-7.08 (m, 4 H), 6.83(d, 1 H), 5.42 (dd, 1 H), 3.65-3.49 (m, 2 H), 3.25 (dd, 1 H) 231

383 (M + NH₄ ⁺) (400 MHz, CDCl₃): δ 7.92 (d, 1 H), 7.21-7.20 (m, 1 H),7.12-7.11 (m, 1 H), 7.03-6.98 (m, 2 H), 5.71-5.65 (m, 1 H), 5.46-5.33(m, 1 H), 3.66 (dd, 1 H), 3.31 (s, 3 H), 3.27-3.05 (m, 2 H) 232

383 (M + NH₄ ⁺) (400 MHz, CDCl₃): δ 7.92 (d, 1 H), 7.21-7.20 (m, 1 H),7.12-7.11 (m, 1 H), 7.03-6.98 (m, 2 H), 5.17-5.65 (m, 1 H), 5.46-5.33(m, 1 H), 3.66 (dd, 1 H), 3.31 (s, 3 H), 3.27-3.05 (m, 2 H) 233

429 (M − H) (400 MHz, CDCl₃): δ 7.88 (d, 1 H), 7.32-7.25 (m, 1 H),7.03-6.98 (m, 1 H), 6.91-6.86 (m, 2 H), 5.42 (dd, 1 H), 3.64-3.47 (m, 2H), 3.22 (d, 1 H) 234

444 (M + HCO₂ ⁻) (400 MHz, CDCl₃): δ 7.92 (d, 1 H), 7.52-7.51 (m, 1 H),7.32-7.31 (m, 1 H), 7.25-7.24 (m, 1 H), 6.98 (d, 1 H), 5.62-5.58 (m, 1H), 3.56-3.35 (m, 3 H), 3.24 (s, 3 H) 235

419 (M − H) (400 MHz, CDCl₃): δ 8.84 (d, 1 H), 8.73 (d, 1 H), 7.96 (d, 1H), 7.75-7.74 (m, 1 H), 6.95 (d, 1 H), 5.45 (dd, 1 H), 3.64-3.48 (m, 2H), 3.31 (d, 1 H) 236

419 (M + NH4) (400 MHz, CDCl₃): δ 7.89 (d, 1H), 7.62-7.57 (m, 2H), 7.42(s, 1H), 7.39- 7.34 (m, 1H), 6.90 (d, 1H), 6.44 (t, 1H), 5.51 (dd, 1H),5.63-5.45 (m, 2H), 3.37 (d, 1H) 237

238

[M + H] 435 (400 MHz, CDCl₃): δ 10.35 (br s, 1H), 8.14 (s, 1H), 7.82 (d,1H), 7.61 (d, 1H), 7.51 (d, 1H), 7.21-7.17 (m, 1H), 6.82 (d, 1H), 5.44(d, 1H), 3.70- 3.57 (m, 2H), 3.40 (br s, 1H) 239

[M + formic acid] 459 (400 MHz, CDCl₃): δ 7.78 (d, 1H), 7.68 (d, 1H),7.44 (d, 1H), 7.37 (t, 1H), 7.23 (d, 1H), 7.00 (d, 1H), 6.71 (d, 1H),5.73-5.68 (m, 1H), 5.38-5.12 (m, 2H), 3.37-3.33 (m, 1H), 3.32-3.22 (m,1H), 3.07-2.99 (m, 1H), 2.56-2.46 (m, 1H), 2.35-2.24 (m, 1H) 240

[M + H] 408 (400 MHz, CD₃OD): δ 8.01 (d, 1H), 7.54-7.49 (m, 1H),7.46-7.44 (m, 1H), 7.40-7.36 (m, 1H), 7.20-7.14 (m, 1H), 5.56 (d, 1H),3.78-3.61 (m, 1H), 3.62 (s, 3H), 3.55-3.47 (m, 1H) 241

[M + 1] 453 (400 MHz, CDCl₃): δ 10.55 (br s, 1H), 7.94 (d, 1H), 7.83 (d,1H), 7.16- 7.10 (m, 1H), 6.86 (d, 1H), 6.83-6.78 (m, 1H), 5.46 (d, 1H),3.72-3.59 (m, 2H), 3.34 (br s, 1H) 242

[M + formic acid] 459 (400 MHz, CDCl₃): δ 7.83-7.76 (m, 2H), 7.47 (d,1H), 7.40 (t, 1H), 7.21- 7.19 (m, 1H), 7.05 (d, 1H), 6.76 (d, 1H),5.61-5.11 (m, 3H), 3.71-3.57 (m, 2H), 3.30 (br s, 1H) 243

[M + H] 435 (400 MHz, CDCl₃): δ 10.45 (br s, 1H), 7.93 (s, 1H), 7.84 (d,1H), 7.48- 7.41 (m, 2H), 6.92 (d, 1H), 6.86 (dd, 1H), 5.46 (d, 1H),3.72-3.59 (m, 2H), 3.44 (br s, 1H) 244

[M + H] 453 (400 MHz, CDCl₃): δ 10.35 (br s, 1H), 7.96 (s, 1H), 7.84 (d,1H), 7.47- 7.43 (m, 1H), 7.39-7.33 (m, 1H), 6.81 (dd, 1H), 5.46 (d, 1H),3.74-3.59 (m, 2H), 3.36 (br s, 1H) 245

[M + H] 408 (400 MHz, CDCl₃): δ 8.01 (d, 1H), 7.32-7.28 (m, 1H),7.22-7.20 (m, 1H), 7.14-7.09 (m, 1H), 7.02 (d, 1H), 5.67 (d, 1H), 4.22(br s, 1H), 3.65 (s, 3H), 3.60-3.40 (m, 2H) 246

[M + H] 435 (400 MHz, CDCl₃): δ 8.15 (d, 1H), 7.82 (d, 1H), 7.72 (s,1H), 7.65 (s, 1H), 6.99 (d, 1H), 6.88-6.81 (m, 2H), 5.43 (d, 1H),3.76-3.63 (m, 2H), 3.51 (br s, 1H) 247

(M + H) 385 (400 MHz, CDCl₃): δ 7.93 (d, 1H), 7.27-7.23 (m, 1H),7.16-7.13 (m, 1H), 7.07-6.98 (m, 2H), 3.56-3.34 (m, 3H), 3.24 (s, 3H)248

(M − H) 402, 404 (400 MHz, CDCl₃): δ 7.70 (d, 1H), 7.20-7.15 (m, 1H),7.10-7.08 (m, 1H), 7.02 (dt, 1H), 6.86 (d, 1H), 3.50 (t, 2H) 249

(M + NH₄) 397 (400 MHz, CDCl₃): δ 7.83 (d, 1H), 7.60 (s, 1H), 7.41 (s,1H), 7.37 (s, 1H), 6.94 (d, 1H), 6.65 (t, 1H), 5.72- 5.68 (m, 1H), 3.64(br d, 1H), 3.22 (s, 3H), 3.14-3.04 (m, 1H), 2.81 (ddd, 1H), 2.54-2.43(m, 1H), 2.28-2.19 (m, 1H) 250

414 (M + H) (400 MHz, CDCl₃): δ 7.90 (d, 1 H), 7.27-7.24 (m, 1 H), 7.16(br s, 1 H), 7.06 (d, 1 H), 7.00 (d, 1 H), 5.62 (d, 1 H), 4.00-4.16 (m,2 H), 3.30-3.74 (m 4 H) 251

392 (M + H) (400 MHz, CDCl₃): δ 7.82 (d, 1H), 6.97 (d, 1H), 6.69 (t,1H), 6.64-6.54 (m, 2H), 5.62 (d, 1H), 5.04-4.96 (m, 1H), 3.50-3.30 (m,2H), 2.64 (d, 3H) 252

383 (M − H) (400 MHz, CDCl₃): δ 7.93 (d, 1H), 7.26-7.20 (m, 1H), 7.12(br s, 1H), 7.04-6.96 (m, 2H), 5.74-5.66 (m, 1H), 5.28 (br s, 2H),3.50-3.32 (m, 2H) 253

368 (M + H) (400 MHz, CDCl₃): δ 8.67 (s, 1H), 8.58 (s, 1H), 7.85 (d,1H), 7.58 (s, 1H), 6.89 (d, 1H), 5.54 (d, 2H), 3.50- 3.24 (m, 2H) 254

399 (M + H) (400 MHz, CDCl₃): δ 7.85 (d, 1H), 7.25-7.18 (m, 1H), 7.13(brs, 1H), 7.08-6.92 (m, 2H), 5.68-5.56 (m, 1H), 5.05 (br s, 1H),3.58-3.30 (m, 2H), 2.65 (s, 3H) 255

322 (M − H) (400 MHz, CDCl₃): δ 8.06 (d, 1H), 6.97-6.93 (m, 1H),6.85-6.83 (m, 1H), 6.69-6.66 (m, 1H), 3.37 (d, 1H), 3.20- 3.12 (m, 1H),2.93-2.85 (m, 1H), 2.52-2.43 (m, 1H), 2.32-2.25 (m, 1H) 256

378 (M + H) (400 MHz, CDCl₃): δ 7.89 (d, 1H), 6.98 (d, 1H), 6.72-6.60(m, 1H), 6.62- 6.52 (m, 2H), 5.72-5.64 (m, 1H), 5.29 (br s, 2H),3.56-3.34 (m, 2H) 257

340 (M − H) (400 MHz, CDCl₃): δ 7.82 (d, 1H), 6.95 (d, 1H), 6.62 (t,1H), 6.55-6.50 (m, 2H), 5.84-5.80 (m, 1H), 5.34 (br s, 2H), 3.11-3.03(m, 1H), 2.83-2.75 (m, 1H), 2.61-2.52 (m, 1H), 2.19-2.10 (m, 1H) 258

(M − H) 399 (400 MHz, CDCl₃): δ 7.96 (d, 1H), 7.27-7.22 (m, 1H),7.18-7.15 (m, 1H), 7.07-7.03 (m, 1H), 6.97 (d, 1H), 5.59 (d, 1H), 4.59(s, 1H), 3.89 (s, 1H), 3.18 (dt, 1H), 2.96 (ddd, 1H), 2.43- 2.27 (m, 2H)259

(M − H) 399 (400 MHz, CDCl₃): δ 8.01 (d, 1H), 7.25-7.21 (m, 1H),7.17-7.14 (m, 1H), 7.06-7.01 (m, 1H), 6.96 (d, 1H), 5.78- 5.73 (m, 1H),3.96-3.93 (m, 1H), 3.73 (s, 1H), 3.13 (dt, 1H), 2.87 (ddd, 1H),2.52-2.41 (m, 1H), 2.31-2.23 (m, 1H) 260

(M + H) 446, 448 (400 MHz, CDCl₃): δ 8.17 (s, 1H), 8.03 (s, 1H), 7.99(d, 1H), 7.11 (d, 1H), 7.08 (s, 1H), 5.44 (dd, 1H), 3.64- 3.42 (m, 3H)261

(M + H) 471 (400 MHz, CDCl₃): δ 7.95-7.87 (m, 2H), 6.95 (d, 1H), 6.77(dd, 1H), 5.46 (d, 1H), 3.66-3.58 (m, 2H), 3.25 (m, 1H) 262

(M + H) 453 (400 MHz, CDCl₃): δ 7.91-7.88 (m, 2H), 7.12 (d, 1H), 7.03(d, 1H), 6.66 (d, 1H), 6.46 (d, 1H), 3.66-3.56 (m, 2H), 3.26 (br s, 1H)263

(M + H) 435 (400 MHz, CDCl₃): δ 7.92 (s, 1H), 7.84 (d, 1H), 7.48-7.42(m, 2H), 6.92 (d, 1H), 6.86 (d, 1H), 5.46 (d, 1H), 3.68-3.59 (m, 2H),3.28 (br s, 1H) 264

(M + H) 453 (400 MHz, CDCl₃): δ 7.93 (d, 1H), 7.84 (d, 1H), 7.13 (t,1H), 6.81 (d, 1H), 6.86 (d, 1H), 5.46 (d, 1H), 3.68- 3.69 (m, 2H), 3.29(br s, 1H) 265

419 (M + H) (400 MHz, CDCl₃): δ 7.90 (d, 1 H), 7.30-7.28 (m, 1 H), 7.19(br s, 1 H), 7.10-7.06 (m, 1 H), 6.92 (d, 1 H), 5.44-5.26 (m, 1 H), 4.93(t, 1 H), 3.40-3.24 (m, 2 H), 1.95 (br s, 2H) 266

(M + H) 453 (400 MHz, CDCl₃): δ 7.96 (s, 1H), 7.83 (d, 1H), 7.36 (t,1H), 6.81 (d, 1H), 6.68 (d, 1H), 5.47 (d, 1H), 3.74- 3.65 (m, 2H), 3.28(br s, 1H) 267

(M − H) 435 (400 MHz, CDCl₃): δ 8.32 (s, 1H), 8.23 (s, 1H), 8.03 (d,1H), 7.23 (s, 1H), 7.13 (d, 1H), 5.46 (dd, 1H), 3.64-3.42 (m, 2H), 3.25(d, 1H) 268

419 (M + H) (400 MHz, CDCl₃): δ 7.90 (d, 1 H), 7.30-7.28 (m, 1 H), 7.22(br s, 1 H), 7.12-7.08 (m, 1 H), 6.95 (d, 1 H), 5.25-5.12 (m, 1 H), 4.95(d, 1 H), 3.52-3.46 (m, 1 H), 3.29-3.18 (m, 1H), 1.73 (br s, 2H) 269

398/400 (M + NH₄ ⁺) (400 MHz, CDCl₃): δ 8.23-8.21 (m, 1 H), 7.35-7.32(m, 1 H), 7.26-7.24 (m, 1 H), 7.23-7.21 (m, 1 H), 7.14-7.10 (m, 1 H),3.97-3.78 (m, 2 H), 3.43 (s, 3 H) 270

417/419 (M + NH₄ ⁺) (400 MHz, CDCl₃): δ 7.95-7.91 (m, 1 H), 7.26-7.23(m, 1 H), 7.14-7.13 (m, 1 H), 7.06-7.00 (m, 2 H), 5.80-5.78 (m, 0.5 H),5.65-5.61 (m, 0.5 H), 3.81-3.55 (m, 3.5 H), 3.25 (s, 1.5 H), 3.24 (s,1.5 H) 271

383 (M + NH₄ ⁺) (400 MHz, CDCl₃): δ 7.87 (d, 1 H), 7.23-7.21 (m, 1 H),7.13-7.12 (m, 1 H), 7.05-7.00 (m, 2 H), 5.62-5.56 (m, 1 H), 5.44-5.29(m, 1 H), 3.66 (dd, 1 H), 3.49-3.35 (m, 1 H), 3.20 (s, 3 H), 3.17-3.06(m, 1 H) 272

[M + H] 368 (400 MHz, CDCl₃): δ 7.84-7.80 (m, 1H), 7.19-7.16 (m, 1H),7.10 (d, 1H), 7.08-7.06 (m, 1H), 7.00-6.96 (m, 1H), 5.40 (d, 1H),4.48-4.36 (m, 1H), 3.49- 3.27 (m, 2H), 2.93 (s, 3H) 273

[M + formate- H] 418 (400 MHz, CDCl₃): δ 7.63 (d, 1H), 7.21-7.18 (m,1H), 7.11-7.09 (m, 1H), 7.03-6.97 (m, 2H), 5.29 (d, 1H), 3.51- 3.28 (m,2H), 2.76 (br s, 1H) 274

(M + H) 401 (400 MHz, CDCl₃): δ 7.95 (d, 1H), 7.28-7.25 (m, 1H),7.17-7.15 (m, 1H), 7.06 (dt, 1H), 7.00 (d, 1H), 5.62-5.56 (m, 1H), 5.37(dd, 1H), 5.24 (dd, 1H), 4.26 (d, 1H), 3.57-3.34 (m, 2H), 3.20 (br d,1H) 275

(M + H) 401 (400 MHz, CDCl₃): δ 7.96 (d, 1H), 7.28-7.25 (m, 1H),7.18-7.16 (m, 1H), 7.06 (dt, 1H), 7.01 (d, 1H), 5.42 (dd, 1H), 5.27 (dd,1H), 5.15 (dd, 1H), 5.04-5.02 (m, 1H), 3.62-3.38 (m, 2H), 3.33 (br s,1H) 276

(M + H) 408 (400 MHz, CDCl₃): δ 8.00 (d, 1H), 7.30 (ddd, 1H), 7.21-7.19(m, 1H), 7.09 (dt, 1H), 6.99 (d, 1H), 5.92 (dd, 1H), 5.76-5.69 (m, 1H),5.65 (dd, 1H), 5.55-5.37 (m, 1H), 3.43-3.18 (m, 2H), 3.22 (dd, 1H) 277

(M + H) 408 (400 MHz, CDCl₃): δ 8.04 (d, 1H), 7.31 (ddd, 1H), 7.24-7.22(m, 1H), 7.11 (dt, 1H), 7.00 (d, 1H), 6.27 (dd, 1H), 5.75-5.69 (m, 1H),5.55 (dd, 1H), 5.56-5.39 (m, 1H), 3.45-3.22 (m, 2H), 3.12 (t, 1H) 278

(M + Cl⁻) 551, 553 (400 MHz, CDCl₃): δ 7.97 (d, 1H), 7.27-7.23 (m, 1H),7.17-7.13 (m, 1H), 7.07 (dt, 1H), 7.02 (d, 1H), 5.54 (dd, 1H), 5.49-5.41(m, 1H), 5.12 (br d, 1H), 3.33 (br s, 1H), 2.73-2.64 (m, 1H), 2.22 (d,1H), 1.35 (s, 9H) 279

(M + H) 417 (400 MHz, CDCl₃): δ 7.93 (d, 1H), 7.31 (ddd, 1H), 7.26-7.24(m, 1H), 7.16 (dt, 1H), 6.94 (d, 1H), 5.53 (d, 1H), 4.59 (d, 1H),2.69-2.61 (m, 1H), 2.35-1.95 (m, 4H) 280

(M + H) 369, 371 (400 MHz, CDCl₃): δ 7.79 (d, 1H), 6.98 (ddd, 1H), 6.91(d, 1H), 6.91- 6.89 (m, 1H), 6.74 (dt, 1H), 5.97 (t, 1H), 5.68 (dt, 1H),5.41 (t, 1H), 3.75 (d, 1H), 3.26-3.17 (m, 1H), 3.20 (s, 3H), 2.91-2.84(m, 1H) 281

(M + H) 387, 389 (400 MHz, CDCl₃): δ 7.85 (d, 1H), 6.95 (ddd, 1H), 6.92(d, 1H), 6.88- 6.85 (m, 1H), 6.70 (dt, 1H), 5.65 (d, 1H), 4.24-4.06 (btm, 1H), 4.08 (dd, 1H), 3.88 (dd, 1H), 3.64-3.59 (m, 1H), 3.26 (s, 3H),2.69 (ddd, 1H), 2.66-2.48 (br m, 1H), 2.12 (d, 1H) 282

(M + H) 390 (400 MHz, CDCl₃): δ 7.93 (d, 1H), 7.27 (ddd, 1H), 7.20-7.18(m, 1H), 7.08 (dt, 1H), 7.00 (d, 1H), 5.98 (dd, 1H), 5.80-5.75 (m, 1H),5.49 (dd, 1H), 3.17 (dt, 1H), 2.94 (ddd, 1H), 2.86 (d, 1H), 2.62-2.51(m, 1H), 2.29- 2.20 (m, 1H) 283

(M + H) 390 (400 MHz, CDCl₃): δ 7.91 (d, 1H), 7.27 (ddd, 1H), 7.19-7.16(m, 1H), 7.06 (dt, 1H), 6.98 (d, 1H), 5.85-5.79 (m, 1H), 5.72 (dd, 1H),5.61 (dd, 1H), 3.15 (ddd, 1H), 2.97 (d, 1H), 2.89 (ddd, 1H), 2.62-2.52(m, 1H), 2.27- 2.18 (m, 1H) 284

(M + H) 479 (400 MHz, CDCl₃): δ 8.04 (d, 1H), 7.30 (ddd, 1H), 7.22-7.19(m, 1H), 7.10 (dt, 1H), 7.01 (d, 1H), 5.97 (dd, 1H), 5.70 (dd, 1H), 5.60(dd, 1H), 3.68 (d, 1H), 3.61-3.39 (m, 2H), 3.23 (s, 3H) 285

(M + H) 426 (400 MHz, CDCl₃): δ 8.01 (d, 1H), 7.34 (ddd, 1H), 7.25-7.22(m, 1H), 7.12 (dt, 1H), 7.01 (d, 1H), 5.75 (dd, 1H), 5.71-5.65 (m, 1H),5.61 (dd, 1H), 3.64-3.45 (m, 2H), 3.14 (dd, 1H) 286

(M + H) 426 (400 MHz, CDCl₃): δ 8.03 (d, 1H), 7.34 (ddd, 1H), 7.26-7.24(m, 1H), 7.14 (dt, 1H), 7.02 (d, 1H), 6.02 (dd, 1H), 5.65-5.59 (m, 1H),5.54 (dd, 1H), 3.66-3.48 (m, 2H), 3.30 (dd, 1H) 287

392 (M + HCO₂ ⁻) (400 MHz, CDCl₃): δ 7.83 (d, 1H), 7.19-7.16 (m, 1H),7.09-7.07 (m, 1H), 7.01-6.96 (m, 2H), 5.71-5.67 (m, 1H), 3.64 (d, 1H),3.21 (s, 3H), 3.12-3.02 (m, 1H), 2.85-2.75 (m, 1H), 2.52-2.42 (m, 1H),2.27-2.18 (m, 1H) 288

428 (M + HCO₂ ⁻) (400 MHz, CDCl₃): δ 8.08 (d, 1H), 7.29-7.23 (m, 1H),7.19 (brs, 1H), 7.15-7.08 (m, 1H), 7.02 (d, 1H), 5.78- 5.70 (m, 1H),3.89 (d, 1H), 3.23 (s, 3H), 3.17-3.02 (m, 1H), 2.80-2.64 (m, 1H) 289

384 (M + H) (400 MHz, CDCl₃): δ 8.13 (d, 1H), 7.31-7.25 (m, 1H),7.23-7.19 (m, 1H), 7.14-7.09 (m, 1H), 7.04 (d, 1H), 6.09- 5.91 (m, 1H),5.87-5.80 (m, 1H), 5.25-5.05 (m, 1H), 3.32 (s, 3H), 2.95 (d, 1H) 290

437 (M + H) (400 MHz, CDCl₃): δ 7.92 (d, 1H), 7.34-7.30 (m, 1H),7.24-7.22 (m, 1H), 7.14-7.10 (m, 1H), 6.94 (d, 1H), 4.85 (d, 1H),3.65-3.41 (m, 2H) 291

481 (M + H) (400 MHz, CDCl₃): δ 7.95 (d, 1H), 7.35-7.31 (m, 1H),7.24-7.22 (m, 1H), 7.14-7.10 (m, 1H), 6.95 (d, 1H), 4.59 (d, 1H),3.77-3.52 (m, 2H), 3.42 (t, 2H), 3.06 (t, 2H) 292

419 (M + NH₄ ⁺) (400 MHz, CDCl₃): δ 8.14-8.11 (m, 1H), 7.33-7.29 (m,1H), 7.25-7.23 (m, 1H), 7.16-7.12 (m, 1H), 7.05 (d, 1H), 5.91-5.75 (m,1H), 5.71-5.65 (m, 1H), 3.39 (d, 1H), 3.25 (s, 3H) 293

419 (M + NH₄ ⁺) (400 MHz, CDCl₃): δ 8.10-8.07 (m, 1H), 7.32-7.28 (m,1H), 7.23-7.20 (m, 1H), 7.15-7.10 (m, 1H), 7.02 (d, 1H), 6.07-5.90 (m,1H), 5.87-5.80 (m, 1H), 3.95 (d, 1H), 3.26 (s, 3H) 294

384 (M + H) (400 MHz, CDCl₃): δ 8.09-8.06 (m, 1H), 7.27-7.24 (m, 1H),7.19-7.17 (m 1H), 7.10-7.07 (m, 1H), 7.04 (d, 1H), 6.30-6.12 (m, 1H),5.96-5.89 (m, 1H), 5.46-5.27 (m, 1H), 3.53-3.51 (m, 1H), 3.27 (s, 3H)295

383 (M + NH₄ ⁺) (400 MHz, CDCl₃): δ 8.04-8.01 (m, 1H), 7.25-7.22 (m,1H), 7.18-7.16 (m, 1H), 7.11-7.06 (m, 1H), 7.00 (d, 1H), 6.09-5.79 (m,1H), 5.69-5.61 (m, 1H), 3.54 (d, 1H), 3.23 (s, 3H), 2.94-2.80 (m, 1H),2.52-2.41 (m, 1H) 296

399 (M + NH₄ ⁺) (400 MHz, CDCl₃): δ 8.05 (d, 1H), 7.26-7.22 (m, 1H),7.19-7.17 (m, 1H), 7.12-7.07 (m, 1H), 7.05 (d, 1H), 5.76- 5.70 (m, 1H),5.30-5.24 (m, 1H), 5.18-5.01 (m, 1H), 3.29 (s, 3H) 297

426 (M + H) (400 MHz, CDCl₃): δ 8.04 (d, 1H), 7.34-7.30 (m, 1H),7.25-7.22 (m, 1H), 7.23 (t, J = 54 Hz, 1H), 7.14-7.10 (m, 1H), 7.00 (d,1H), 5.71-5.63 (m, 1H), 5.56-5.52 (m, 0.5H), 5.43-5.39 (m, 0.5H), 3.59(t, 1H), 3.46-3.18 (m, 2H) 298

397 (M + NH₄) (400 MHz, CDCl₃): δ 7.85 (d, 1H), 7.22-7.19 (m, 1H),7.12-7.09 (m, 1H), 7.03-6.98 (m, 2H), 5.29-5.23 (m, 1H), 3.57-3.53 (m,1H), 3.26-3.04 (m, 2H), 3.19 (s, 3H), 1.70 (d, J = 22 Hz, 3H) 299

426 (M + H) (400 MHz, CDCl₃): δ 8.03 (d, 1H), 7.34-7.30 (m, 1H),7.23-7.21 (m, 1H), 7.13-7.09 (m, 1H), 7.01 (t, J = 53 Hz, 1H), 6.99 (d,1H), 5.73-5.66 (m, 1H), 5.56-5.52 (m, 0.5H), 5.43-5.39 (m, 0.5H),3.45-3.34 (m, 1H), 3.35-3.19 (m, 2H) 300

397 (M + NH₄) (400 MHz, CDCl₃): δ 7.90 (d, 1H), 7.21-7.19 (m, 1H),7.10-7.08 (m, 1H), 6.99 (dt, 1H), 6.98 (d, 1H), 5.40-5.35 (m, 1H),3.79-3.77 (m, 1H), 3.36-3.27 (m, 1H), 3.32 (s, 3H), 2.95-2.84 (m, 1H),1.70 (d, 3H) 301

379 (M + NH₄) (400 MHz, CDCl₃): δ 7.84 (d, 1H), 7.19-7.15 (m, 1H),7.07-7.06 (m, 1H), 6.98 (d, 1H), 6.97 (dt, 1H), 5.46-5.43 (m, 1H), 3.12(s, 3H), 3.08 (d, 1H), 2.97-2.91 (m, 1H), 2.68-2.53 (m, 2H), 1.25 (d,3H) 302

390 (M + H) (400 MHz, CDCl₃): δ 8.00 (d, J = 8.7 Hz, 0.5H), 7.95 (d, J =8.7 Hz, 0.5H), 7.29-7.25 (m, 1H), 7.19-7.16 (m, 1H), 7.10-7.05 (m, 1H),7.01 (d, 1H), 5.78- 5.69 (m, 1H), 5.54-5.50 (m, 0.5H), 5.40-5.37 (m,0.5H), 3.50 (d, J = 42 Hz, 3H), 3.39-3.11 (m, 3H) 303

[M + H] = 357 (400 MHz, CDCl₃): δ 8.72 (s, 1H), 8.64 (s, 1H), 7.64 (d,1H), 7.59-7.57 (m, 1H), 6.97 (d, 1H), 5.33-5.28 (m, 1H), 3.55-3.32 (m,2H), 2.86-2.82 (m, 1H) 304

437, 439 (M + H⁺) (400 MHz, CDCl₃): δ 7.88 (d, 1 H), 7.17-7.13 (m, 1 H),7.04-7.02 (m, 1 H), 6.98 (d, 1 H), 6.77-6.74 (m, 1 H), 5.61-5.56 (m, 1H), 3.57-3.36 (m 3 H), 3.22 (s, 3 H) 305

(M + H) 437 (400 MHz, CDCl₃): δ 8.01 (d, 1H), 7.32 (ddd, 1H), 7.24-7.22(m, 1H), 7.12 (dt, 1H), 6.99 (d, 1H), 5.35 (dd, 1H), 4.73-4.71 (m, 1H),3.97 (br s, 1H), 3.63-3.46 (m, 2H) 306

(M + H) 437 (400 MHz, CDCl₃): δ 8.07 (d, 1H), 7.30 (ddd, 1H), 7.23-7.21(m, 1H), 7.11 (dt, 1H), 6.98 (d, 1H), 5.59 (ddd, 1H), 3.97 (d, 1H), 3.81(br s, 1H), 3.61-3.39 (m, 2H) 307

(M + H) 477 (400 MHz, CDCl₃): δ 7.99 (d, 1H), 7.30 (ddd, 1H), 7.23-7.20(m, 1H), 7.10 (dt, 1H), 6.98 (d, 1H), 6.04-5.93 (m, 1H), 5.35-5.28 (m,2H), 5.21 (dq, 1H), 4.87 (br s, 1H), 4.16-4.09 (m, 1H), 4.04-3.96 (m,1H), 3.61-3.44 (m, 2H) 308

(M + H) 477 (400 MHz, CDCl₃): δ 8.04 (d, 1H), 7.29 (ddd, 1H), 7.21-7.19(m, 1H), 7.09 (dt, 1H), 6.97 (d, 1H), 5.98 (ddt, 1H), 5.58 (dd, 1H),5.34 (dq, 1H), 5.19 (dq, 1H), 4.13-4.05 (m, 1H), 4.03-3.95 (m, 1H),3.59-3.33 (m, 3H) 309

(M + H) 414 (400 MHz, CDCl₃): δ 8.49 (d, 1H), 8.37 (d, 1H), 7.93 (d,1H), 7.26 (dt, 1H), 6.95 (d, 1H), 5.44 (dd, 1H), 3.67-3.48 (m, 2H), 3.42(d, 1H). 310

(M + H) 430 (400 MHz, CDCl₃): δ 8.11-8.08 (m, 1H), 8.01-7.97 (m, 2H),7.14 (d, 1H), 6.90 (dt, 1H), 5.43 (dd, 1H), 3.95 (d, 1H), 3.62-3.41 (m,2H) 311

400 (M + H) (400 MHz, CDCl₃): δ 8.00 (d, 1H), 7.27-7.25 (m, 1H),7.20-7.18 (m, 1H), 7.12-7.07 (m, 1H), 7.03 (d, 1H), 5.81- 5.74 (m, 1H),5.43-5.36 (m, 1H), 3.81 (d, 1H), 3.25 (s, 3H), 2.71 (m, 1H) 312

400 (M + H) (400 MHz, CDCl₃): δ 8.05 (d, 1H), 7.29-7.20 (m, 2H),7.15-7.10 (m, 1H), 7.05 (d, 1H), 5.63-5.57 (m, 1H), 5.22- 5.15 (m, 1H),3.53-3.48 (m, 1H), 3.24 (s, 3H), 2.73 (d, 1H) 313

349 (M + H) (400 MHz, CDCl₃): δ 8.75-8.72 (m, 1H), 8.66-8.64 (m, 1H),7.92 (d, 1H), 7.61-7.59 (m, 1H), 6.95 (d, 1H), 5.73- 5.65 (m, 1H),5.51-5.47 (m, 0.5H), 5.38-5.34 (m, 0.5H), 3.71-3.68 (m, 1H), 3.36-3.38(m, 2H), 3.31 (s, 3H) 314

(M + H) 420 (400 MHz, CDCl₃): δ 8.83 (d, 1H), 8.72 (d, 1H), 8.02 (d,1H), 7.73 (dd, 1H), 6.95 (d, 1H), 5.35 (dd, 1H), 4.73-4.70 (m s, 1H),3.99 (br s, 1H), 3.67-3.49 (m, 2H) 315

(M + H) 420 (400 MHz, CDCl₃): δ 8.82 (d, 1H), 8.71 (d, 1H), 8.08 (d,1H), 7.72 (dd, 1H), 6.93 (d, 1H), 5.63-5.57 (m, 1H), 3.97 (d, 1H), 3.82(br s, 1H), 3.65- 3.43 (m, 2H) 316

[M + H] 426 (400 MHz, CD₃OD): δ 8.21 (dd, 1H), 7.59-7.56 (m, 1H),7.54-7.53 (m, 1H), 7.46 (dt, 1H), 7.25 (d, 1H), 6.00 (dd, 1H), 5.60-5.56(m, 1H), 3.64 (s, 3H) 317

[M + H] 426 (400 MHz, CDCl₃): δ 8.23-8.20 (m, 1H), 7.38-7.34 (m, 1H),7.30-7.28 (m, 1H), 7.21-7.17 (m, 1H), 7.09 (d, 1H), 5.90 (dd, 1H),5.71-5.66 (m, 1H), 3.90-3.88 (m, 1H), 3.64 (s, 3H) 318

[M + H] 426 (400 MHz, CDCl₃): δ 8.13 (dd, 1H), 7.37-7.33 (m, 1H),7.28-7.27 (m, 1H), 7.19-7.15 (m, 1H), 7.05 (d, 1H), 6.08-5.84 (m, 2H),4.08 (d, 1H), 3.54 (s, 3H) 319

[M + H] 409 (400 MHz, CD₃COCD₃): δ 8.95 (d, 1H), 8.94 (d, 1H), 8.34-8.32(m, 1H), 8.23-8.20 (m, 1H), 7.45 (d, 1H), 6.43- 6.40 (m, 1H), 6.15 (dd,1H), 5.72-5.66 (m, 1H), 3.69 (s, 3H) 320

[M + H] 409 (400 MHz, CD₃COCD₃): δ 8.96-8.95 (m, 1H), 8.94-8.92 (m, 1H),8.34-8.32 (m, 1H), 8.24-8.20 (m, 1H), 7.44-7.41 (m, 1H), 6.51-6.31 (m,2H), 5.90-5.83 (m, 1H), 3.81 (s, 3H) 321

[M + H] 409 (400 MHz, CD₃COCD₃): δ 8.97-8.96 (m, 1H), 8.96-8.94 (m, 1H),8.35 (dd, 1H), 8.25 (dd, 1H), 7.45 (d, 1H), 6.50 (brs, 1H), 6.16 (dd,1H), 5.68-5.30 (m, 1H), 3.80 (s, 3H) 322

[M + H] 409 (400 MHz, CD₃COCD₃): δ 8.95-8.94 (m, 1H), 8.94-8.92 (m, 1H),8.33 (dd, 1H), 8.19 (dd, 1H), 7.41 (d, 1H), 6.50-6.28 (m, 2H), 5.90-5.85(m, 1H), 3.70 (s, 3H) 323

418, 420 (M + H) (400 MHz, CD₃COCD₃): δ 8.65-8.63 (m, 1H), 8.60-8.59 (m,1H), 8.15-8.12 (m, 1H), 8.02-8.00 (m, 1H), 7.60-7.57 (m, 1H), 3.86-3.79(m, 2H), 3.39 (s, 3H) 324

420, 422 (M + H) (400 MHz, CDCl₃): δ 8.61-8.59 (m, 1H), 8.41-8.39 (m,1H), 7.91-7.87 (d, 1H), 7.61-7.58 (m, 1H), 6.95-6.9′ (d, 1H), 5.63-5.57(m, 1H), 3.61-3.40 (m, 3H), 3.23 (s, 3H) 325

367 (M + H) (400 MHz, CDCl₃): δ 8.76-8.75 (m, 1H), 8.67-8.66 (m, 1H),7.95 (d, 1H), 7.70-7.68 (m, 1H), 6.98 (d, 1H), 5.60 (d, 1H), 3.57-3.35(m, 3H), 3.22 (s, 3H) 326

402, 404 (M + H) (400 MHz, CDCl₃): δ 8.57-8.56 (m, 1H), 8.39-8.38 (m,1H), 7.88 (d, 1H), 7.56-7.54 (m, 1H), 6.91 (d, 1H), 5.72- 5.65 (m, 1H),5.51-5.47 (m, 0.5H), 5.38-5.34 (m, 0.5H), 3.71-3.69 (m, 1H), 3.38-3.09(m, 3H), 3.29 (s, 3H) 327

402 (M + H) (400 MHz, CDCl₃): δ 8.10-8.06 (m, 1H), 7.44-7.32 (m, 3H),6.91 (d, 1H), 5.95-5.91 (m, 0.5H), 5.81-5.78 (m, 0.5H), 5.70-5.64 (m,1H), 4.00-3.97 (m, 1H), 3.24 (s, 3H) 328

420, 422 (M + H) (400 MHz, CDCl₃): δ 8.63-8.61 (m, 1H), 8.45-8.43 (m,1H), 8.11-8.07 (m, 1H), 7.66-7.64 (m, 1H), 6.96 (d, 1H), 6.13-6.11 (m,0.5H), 5.99-5.97 (m, 0.5H), 5.86-5.82 (m, 1H), 5.24-5.04 (m, 1H), 3.30(s, 3H), 3.03-3.00 (m, 1H) 329

378 (M + H) (400 MHz, CDCl₃): δ 8.49-8.47 (m, 1H), 8.39-8.37 (m, 1H),8.11-8.07 (m, 1H), 7.29-7.25 (m, 1H), 7.00 (d, 1H), 5.96-5.93 (m, 0.5H),5.83-5.79 (m, 0.5H), 5.71-5.65 (m, 1H), 3.65-3.63 (m, 1H), 3.24 (s, 3H)330

378 (M + H) (400 MHz, CDCl₃): δ 8.49-8.46 (m, 1H), 8.39-8.36 (m, 1H),8.08-8.04 (m, 1H), 8.28-8.24 (m, 1H), 6.98 (d, 1H), 6.12-6.08 (m, 0.5H),5.99-5.95 (m, 0.5H), 5.88-5.81 (m, 1H), 4.10-4.06 (m, 1H), 3.26 (s, 3H)331

394, 396 (M + H) (400 MHz, CDCl₃): δ 8.56-8.55 (m, 1H), 8.44-8.43 (m,1H), 8.11-8.08 (m, 1H), 7.54-7.52 (m, 1H), 6.99 (d, 1H), 5.95-5.92 (m,0.5H), 5.83-5.79 (m, 0.5H), 5.71-5.65 (m, 1H), 3.66-3.64 (m, 1H), 3.25(s, 3H) 332

394, 396 (M + H) (400 MHz, CDCl₃): δ 8.56-8.54 (m, 1H), 8.43-8.41 (m,1H), 8.08-8.04 (m, 1H), 7.52-7.50 (m, 1H), 6.96 (d, 1H), 6.12-6.08 (m,0.5H), 5.98-5.94 (m, 0.5H), 5.88-5.81 (m, 1H), 4.02-3.99 (m, 1H), 3.26(s, 3H) 333

367 (M + H) (400 MHz, CD₃COCD₃): δ 8.88-8.86 (m, 1H), 8.82-8.80 (m, 1H),8.13-8.08 (m, 2H), 7.33 (d, 1H), 6.21-6.18 (m, 0.5H), 6.07-6.04 (m,0.5H), 5.83-5.79 (m, 1H), 5.36-5.29 (m, 0.5H), 5.25- 5.16 (m, 0.5H),5.07-5.04 (m, 1H), 3.33 (s, 3H) 334

365 (M − H) (400 MHz, CD₃OD): δ 7.87 (d, 1H), 7.42-7.35 (m, 1H),7.26-7.13 (m, 2H), 7.08 (d, 1H), 5.63-5.51 (m, 1H), 5.40- 5.18 (m, 1H),3.20-3.15 (m, 2H) 335

383 (M − H) (400 MHz, CD₃OD): δ 8.04 (d, 1H), 7.45-7.41 (m, 1H),7.31-7.29 (m, 1H), 7.26-7.21 (m, 1H), 7.18 (d, 1H), 6.30- 6.11 (m, 1H),5.80 (t, 1H), 5.37-5.17 (m, 1H) 336

383 (M − H) 337

399 (M − H) (400 MHz, CD₃OD): δ 8.00 (d, 1H), 7.44-7.41 (m, 1H),7.35-7.32 (m, 1H), 7.29-7.24 (m, 1H), 7.14 (d, 1H), 5.46 (d, 1H), 5.06(d, 1H) 338

401 (M − H) (400 MHz, CD₃OD): δ 8.03-8.00 (m, 1H), 7.24-7.20 (m, 1H),7.17-7.15 (m, 1H), 7.08-7.04 (m, 1H), 6.96 (d, 1H), 5.82-5.65 (m, 1H),5.54-5.48 (m, 1H) 339

401 (M − H) (400 MHz, CD₃OD): δ 8.09-8.05 (m, 1H), 7.50-7.46 (m, 1H),7.39-7.38 (m, 1H), 7.33-7.29 (m, 1H), 7.14 (d, 1H), 6.19-6.02 (m, 1H),5.72-5.65 (m, 1H) 340

384 (M − H) (400 MHz, CD₃OD): δ 8.81 (d, 1H), 8.73 (d, 1H), 8.11-8.07(m, 1H), 8.06- 8.04 (m, 1H), 7.18 (d, 1H), 6.04-5.86 (m, 1H), 5.57-5.51(m, 1H) 341

377 (M − H) (400 MHz, CD₃OD): δ 8.43 (d, 1H), 8.35 (d, 1H), 8.10-8.06(m, 1H), 7.59- 7.54 (m, 1H), 7.15 (d, 1H), 6.03-5.85 (m, 1H), 5.56-5.50(m, 1H) 342

(M + H) 385 (400 MHz, CDCl₃): δ 8.82 (d, 1H), 8.74 (d, 1H), 8.14 (dd,1H), 7.74 (dd, 1H), 7.02 (d, 1H), 5.87 (dd, 1H), 5.73-5.66 (m, 1H), 3.58(d, 1H), 3.26 (s, 3H) 343

(M + H) 385 (400 MHz, (CD₃)₂CO): δ 8.89 (dd, 1H), 8.86 (d, 1H), 8.21(dd, 1H), 8.11 (dd, 1H), 7.36 (d, 1H), 6.36 (ddd, 1H), 6.10 (d, 1H),5.87-5.80 (m, 1H), 3.31 (s, 3H) 344

(M + H) 403 (400 MHz, CDCl₃): δ 8.84 (d, 1H), 8.75 (d, 1H), 8.15 (dd,1H), 7.77 (dd, 1H), 7.02 (d, 1H), 5.83 (dd, 1H), 5.68-5.62 (m, 1H), 5.43(dd, 1H), 5.31 (dd, 1H), 3.43 (dd, 1H) 345

(M + H) 403 (400 MHz, (CD₃)₂CO): δ 8.93 (dd, 1H), 8.90 (dd, 1H), 8.26(dd, 1H), 8.13 (dd, 1H), 7.38 (d, 1H), 6.39 (ddd, 1H), 5.73 (dd, 1H),5.80 (ddd, 1H), 5.61 (dd, 1H) 346

(M + H) 401 (400 MHz, CDCl₃): δ 8.14 (dd, 1H), 7.30 (ddd, 1H), 7.24-7.22(m, 1H), 7.14 (dt, 1H), 6.98 (d, 1H), 5.77 (dd, 1H), 5.10-5.01 (m, 1H),3.45 (s, 3H), 1.82 (br d, 2H) 347

(400 MHz, CDCl₃): δ 7.86 (d, 1H), 7.27 (d, 1H), 7.27-7.24 (m, 1H), 7.13-7.11 (m, 1H), 7.04-7.00 (m, 1H), 5.41-5.37 (m, 1H), 3.06 (d, 1H) 348

352, 354 (M − H) (CDCl₃, 400 MHz) δ 7.89 (d, 1H), 7.13 (d, 1H),7.09-7.06 (m, 1H), 6.96-6.94 (m, 1H), 6.80 (dt, 1H), 5.78-5.74 (m, 1H),3.91 (dd, 1H), 3.65 (dd, 1H), 3.41 (d, 1H) 349

404 (M − H) (400 MHz, CDCl₃): δ 7.98 (d, 1H), 7.33-7.30 (m, 1H), 7.23(t, 1H), 7.22- 7.18 (m, 2H), 7.10-7.06 (m, 1H), 5.69-5.65 (m, 1H), 3.23(d, 1H) 350

391, 393, 395 (M + H) (400 MHz, CDCl₃): δ 7.70 (d, 1H), 7.10 (d, 1H),6.93 (ddd, 1H), 6.78- 6.76 (m, 1H), 6.63 (dt, 1H), 3.62-3.58 (m, 2H),3.42-3.37 (m, 2H) 351

336, 338 (M + H) (400 MHz, CDCl₃): δ 7.87 (d, 1H), 7.06 (dt, 1H), 7.03(d, 1H), 6.94-6.92 (m, 1H), 6.78 (dt, 1H), 3.66-3.61 (m, 2H), 3.60-3.55(m, 2H) 352

424, 426, 428 (M + NH₄) (400 MHz, CDCl₃): δ 7.72 (d, 1H), 7.19 (d, 1H),6.98-6.94 (ddd, 1H), 6.82-6.80 (m, 1H), 6.67 (dt, 1H), 5.60 (td, 1H),3.80 (dd, 1H), 3.68 (dd, 1H), 2.89 (d, 1H) 353

352, 354 (M − H) (400 MHz, CDCl₃): δ 7.89 (d, 1H), 7.13 (d, 1H), 7.07(ddd, 1H), 6.97- 6.94 (m, 1H), 6.80 (dt, 1H), 5.79-5.72 (m, 1H), 3.91(dd, 1H), 3.64 (dd, 1H), 3.57 (br d, 1H) 354

(400 MHz, CDCl₃): δ 7.89 (d, 1H), 7.50 (d, 1H), 7.06 (ddd, 1H), 6.89-6.86 (m, 1H), 6.73 (dt, 1H) 355

425, 427, 429 (M − OH) (400 MHz, CDCl₃): δ 7.80 (d, 1H), 7.22 (d, 1H),7.01 (dt, 1H), 6.87-6.85 (m, 1H), 6.71 (dt, 1H), 5.38 (d, 1H), 2.98 (brs, 1H) 356

388, 390 (M − H) (400 MHz, CDCl₃): δ 7.97 (d, 1H), 7.19 (d, 1H), 7.12(ddd, 1H), 6.99- 6.97 (m, 1H), 6.83 (dt, 1H), 5.58-5.51 (m, 1H), 3.51(br d, 1H) 357

397, 399 (M − H) (400 MHz, CDCl₃): δ 7.76 (d, 1H), 7.26 (d, 1H), 7.01(ddd, 1H), 6.87- 6.85 (m, 1H), 6.71 (dt, 1H), 5.44 (dd, 1H), 2.94 (d,1H) 358

364, 366 (M + H) (400 MHz, CDCl₃): δ 7.81 (d, 1H), 7.36 (dd, 1H), 7.20(ddd, 1H), 6.99 (ddd, 1H), 6.89-6.86 (m, 1H), 6.71 (dt, 1H), 4.73-4.61(m, 1H), 1.79 (br d, 2H) 359

377, 379 (M − H) (400 MHz, CDCl₃): δ 7.70 (d, 1H), 7.16 (d, 1H), 6.95(ddd, 1H), 6.80- 6.78 (m, 1H), 6.64 (dt, 1H), 5.30 (dd, 1H), 2.72 (dd,1H), 2.43 (s, 3H) 360

431, 433 (M − H) (400 MHz, CDCl₃): δ 8.00 (d, 1H), 7.29 (d, 1H), 7.04(ddd, 1H), 6.91- 6.89 (m, 1H), 6.74 (dt, 1H), 5.58 (d, 1H), 3.16 (br s,1H) 361

377, 379 (M − H) (400 MHz, CDCl₃): δ 7.85 (d, 1H), 7.22 (t, 1H), 7.14(dt, 1H), 7.01 (ddd, 1H), 6.87-6.85 (m, 1H), 6.71 (dt, 1H), 5.90-5.85(m, 1H), 3.77 (ddd, 1H), 3.67 (dd, 1H), 2.87 (t, 1H) 362

363, 365 (M − H) (400 MHz, CDCl₃): δ 7.84 (d, 1H), 7.27-7.23 (m, 2H),7.01 (dt, 1H), 6.90-6.88 (m, 1H), 6.72 (dt, 1H), 5.35 (q, 1H), 2.79 (dd,1H) 363

378, 380 (M + H) (400 MHz, CDCl₃): δ 7.80 (d, 1H), 7.22 (t, 1H), 7.06(dt, 1H), 7.00 (dt, 1H), 6.87-6.84 (m, 1H), 6.70 (dt, 1H), 5.16-5.03 (brs, 1H), 3.75 (dd, 1H), 3.49 (dd, 1H), 2.20-1.97 (br s, 2H) 364

413, 415 (M − H) (400 MHz, CDCl₃): δ 7.93 (d, 1H), 7.25 (t, 1H),7.21-7.17 (m, 1H), 7.06 (ddd, 1H), 6.92-6.89 (m, 1H), 6.75 (dt, 1H),5.67 (dd, 1H), 3.10 (dd, 1H) 365

398 (M + H) (400 MHz, CDCl₃): δ 7.90 (d, 1H), 7.28 (dd, 1H), 7.18-7.14(m, 1H), 6.76 (tt, 1H), 6.67-6.60 (m, 2H), 4.98 (dt, 1H), 2.01 (br d,2H) 366

380 (M + H) (400 MHz, CDCl₃): δ 7.87 (d, 1H), 7.44 (dd, 1H), 7.15 (d,1H), 6.73 (tt, 1H), 6.64-6.57 (m, 2H), 5.58 (dd, 1H), 5.17-5.07 (m, 1H),2.02-1.93 (m, 2H) 367

362 (M + H) (400 MHz, CDCl₃): δ 7.81 (d, 1H), 7.22 (t, 1H), 7.10-7.06(m, 1H), 6.72 (tt, 1H), 6.63-6.56 (m, 2H), 5.14-5.07 (m, 1H), 3.75 (dd,1H), 3.49 (dd, 1H), 2.12-2.04 (m, 2H) 368

397 (M − H) (400 MHz, CDCl₃): δ 7.93 (d, 1H), 7.25 (t, 1H), 7.23-7.19(m, 1H), 6.78 (tt, 1H), 6.68-6.61 (m, 2H), 5.67 (dd, 1H), 3.09 (dd, 1H)369

379 (M − H) (400 MHz, CDCl₃): δ 7.89 (d, 1H), 7.29 (t, 1H), 7.19 (d,1H), 6.74 (tt, 1H), 6.65-6.58 (m, 2H), 5.87-5.80 (m, 1H), 5.66 (dd, 1H),2.98 (ddd, 1H) 370

363 (M + H) (400 MHz, CDCl₃): δ 7.85 (d, 1H), 7.25 (t, 1H), 7.16 (dt,1H), 6.73 (tt, 1H), 6.63-6.56 (m, 1H), 5.90-5.86 (m, 1H), 5.90-5.85 (m,1H), 3.78 (ddd, 1H), 3.67 (dd, 1H), 2.89 (t, 1H) 371

414, 416 (M − H) (400 MHz, CDCl₃): δ 7.95-7.92 (m, 1H), 7.25 (t, 1H),7.21-7.17 (m, 1H), 7.06 (ddd, 1H), 6.92-6.89 (m, 1H), 6.75 (dt, 1H),3.07 (d, 1H) 372

405 (M + H) (400 MHz, CDCl₃): δ 7.95 (d, 1H), 7.31-7.27 (m, 1H), 7.30(dd, 1H), 7.19-7.14 (m, 2H), 7.07 (dt, 1H), 5.00-4.92 (m, 1H), 2.03 (d,2H) 373

385 (M + H) (400 MHz, CDCl₃): δ 11.26-11.22 (m, 1H), 8.09 (dd, 1H), 8.06(d, 1H), 7.04 (d, 1H), 7.27-7.23 (m, 1H), 7.15-7.13 (m, 1H), 7.06 (dt,1H), 5.87 (dd, 1H) 374

370 (M − H) (400 MHz, CDCl₃): δ 7.89 (d, 1H), 7.28-7.25 (m, 1H), 7.20(t, 1H), 7.17- 7.13 (m, 2H), 7.04 (dt, 1H), 5.90-5.85 (m, 1H), 3.79 (dd,1H), 3.69 (dd, 1H), 2.93 (t, 1H) 375

404 (M − H) (400 MHz, CDCl₃): δ 7.98 (d, 1H), 7.31 (ddd, 1H), 7.23 (t,1H), 7.22-7.18 (m, 2H), 7.08 (dt, 1H), 5.66 (dd, 1H), 3.23 (d, 1H) 376

426, 428 (M + H) (400 MHz, CDCl₃): δ 7.78 (d, 1H), 7.18 (d, 1H), 6.72(tt, 1H), 6.63-6.54 (m, 2H), 4.70 (dt, 1H), 1.92 (d, 2H) 377

426, 428 (M + H) (400 MHz, CDCl₃): δ 7.88 (d, 1H), 7.22 (d, 1H), 6.71(tt, 1H), 6.62-6.55 (m, 2H), 5.31 (dd, 1H), 3.43 (br s, 1H), 3.03 (d,1H) 378

426, 428 (M + H) (400 MHz, CDCl₃): δ 7.81 (d, 1H), 7.22 (d, 1H), 6.70(tt, 1H), 6.61-6.54 (m, 2H), 5.35 (t, 1H), 3.75 (br s, 1H), 3.27 (d, 1H)379

484, 486 (M + H) (400 MHz, CDCl₃): δ 7.75 (d, 1H), 7.16 (d, 1H), 6.71(tt, 1H), 6.62-6.55 (m, 2H), 4.60 (dd, 1H), 3.55 (t, 2H), 3.35 (s, 3H),3.21-3.06 (m, 2H), 2.10- 2.03 (m, 1H) 380

404 (M − H) (400 MHz, CDCl₃): δ 7.98 (d, 1H), 7.31 (ddd, 1H), 7.23 (t,1H), 7.22-7.18 (m, 2H), 7.08 (dt, 1H), 5.66 (dd, 1H), 3.23 (d, 1H) 381

398, 400 (M + H) (400 MHz, CDCl₃): δ 8.55 (d, 1H), 8.38 (d, 1H), 7.96(d, 1H), 7.46 (t, 1H), 7.29 (t, 1H), 7.16-7.13 (m, 1H), 5.67 (dd, 1H),3.24 (dd, 1H) 382

463 (M + H) (400 MHz, CDCl₃): δ 7.91 (d, 1H), 7.39 (t, 1H), 7.29-7.24(m, 1H), 7.19- 7.14 (m, 2H), 7.05 (dt, 1H), 4.94-4.87 (m, 1H), 3.53 (t,2H), 3.37 (s, 3H), 3.17-3.07 (m, 2H), 2.28-2.20 (m, 1H) 383

389 (M + H) (400 MHz, CD₃OD): δ 8.81 (d, 1H), 8.71 (d, 1H), 8.08 (d,1H), 8.04 (dd, 1H), 7.42 (d, 1H), 7.38 (t, 1H), 5.69 (d, 1H) 384

405 (M + H) (400 MHz, CDCl₃): δ 7.95 (d, 1H), 7.31-7.27 (m, 1H), 7.30(dd, 1H), 7.19-7.14 (m, 2H), 7.07 (dt, 1H), 5.01-4.91 (dd, 1H),2.06-1.99 (m, 2H) 385

(400 MHz, (CD₃)₂SO): δ 8.79 (s, 2H), 8.27 (d, 1H), 7.78-7.73 (m, 1H),7.72 (t, 1H), 7.55-7.47 (m, 3H) 386

405 (M − H) (400 MHz, CDCl₃): δ 7.98 (d, 1H), 7.33-7.30 (m, 1H), 7.23(t, 1H), 7.22- 7.18 (m, 2H), 7.08 (dt, 1H), 3.12 (s, 1H) 387

405 (M + H) (400 MHz, CDCl₃): δ 7.94 (d, 1H), 7.30 (d, 1H), 7.31-7.27(m, 1H), 7.19- 7.14 (m, 2H), 7.08 (dt, 1H), 5.02-4.89 (m, 1H), 2.12-1.92(m, 2H) 388

405 (M + NH₄) (400 MHz, CDCl₃): δ 7.90 (d, 1H), 7.29-7.25 (m, 1H), 7.19(d, 1H), 7.16- 7.14 (m, 1H), 7.06-7.01 (m, 1H), 5.75 (d, 2H), 5.58 (brd, 1H), 3.30-3.22 (m, 1H) 389

405 (M − H) (400 MHz, CDCl₃): δ 7.98 (d, 1H), 7.33-7.30 (m, 1H), 7.23(t, 1H), 7.22- 7.18 (m, 2H), 7.08 (dt, 1H), 3.12 (s, 1H) 390

373 (M + NH₄) (400 MHz, CD₃CN): δ 8.02 (d, 1H), 7.27 (d, 1H), 7.01-6.87(m, 3H), 5.88- 5.73 (m, 2H), 4.91 (d, 1H) 391

(400 MHz, CDCl₃): δ 8.13 (d, 1H), 7.67 (t, 1H), 7.53 (d, 1H), 7.43 (s,1H), 7.21 (s, 1H), 7.15 (s, 1H), 2.45 (s, 3H) 392

419 (M + NH₄) (400 MHz, CDCl₃): δ 7.91 (d, 1H), 7.41 (s, 1H), 7.27 (t,1H), 7.21 (s, 1H), 7.13 (s, 1H), 7.10 (d, 1H), 5.71- 5.64 (m, 1H), 3.04(br d, 1H), 2.44 (s, 3H) 393

467, 469 (M + H2O + NH4) (400 MHz, CDCl₃): δ 8.05 (d, 1H), 7.55 (d, 1H),7.30 (d, 1H), 7.13 (s, 1H), 7.07-7.02 (m, 1H) 394

387 (M + H) (400 MHz, CDCl₃): δ 8.01 (d, 1H), 7.28 (t, 1H), 7.21 (d,1H), 6.83-6.76 (m, 1H), 6.69-6.63 (m, 2H), 6.03-5.97 (m, 1H), 4.20 (dd,1H), 3.88 (dd, 1H), 3.15-3.11 (m, 1H) 395

387 (M + H) (400 MHz, CDCl₃): δ 7.97 (d, 1H), 7.26 (t, 1H), 7.21 (d,1H), 6.83-6.76 (m, 1H), 6.70-6.63 (m, 2H), 5.99 (t, 1H), 4.08-4.02 (m,1H), 3.98 (dd, 1H), 3.20-3.15 (m, 1H) 396

362 (M + H) (400 MHz, CDCl³): δ 7.80 (d, 1H), 7.17 (t, 1H), 7.12 (d,1H), 6.74-6.67 (m, 1H), 6.62-6.55 (m, 2H), 5.80-5.74 (m, 1H), 3.80 (dd,1H), 3.75-3.69 (m, 2H), 3.41-3.34 (m, 1H) 397

423 (M + H) (400 MHz, CDCl₃): δ 8.07-8.00 (m, 1H), 7.42-7.13 (m, 2H),6.87-6.80 (m, 1H), 6.73-6.66 (m, 2H), 5.83-5.76 (m, 1H), 3.78-3.66 (m,1H) 398

398 (M + H) (400 MHz, CDCl₃): δ 8.02 (d, 1H), 7.22 (t, 1H), 7.19 (d,1H), 6.79-6.72 (m, 1H), 6.66-6.59 (m, 2H), 5.60 (t, 1H), 3.39 (br s,1H), 3.12 (d, 1H) 399

398 (M + H) (400 MHz, CDCl₃): δ 7.95 (d, 1H), 7.23 (t, 1H), 7.19 (d,1H), 6.79-6.72 (m, 1H), 6.66-6.59 (m, 2H), 5.64 (t, 1H), 3.68 (br s,1H), 3.33 (d, 1H) 400

387 (M + H) (400 MHz, CD₃OD): δ 7.99 (d, 1H), 7.57 (d, 1H), 7.26 (t,1H), 5.57-5.52 (m, 1H), 4.95-4.88 (m, 1H), 3.98-3.91 (m, 2H), 3.67-3.60(m, 2H), 2.32-2.03 (m, 2H), 1.86-1.76 (m, 2H) 401

398 (M + H) (400 MHz, CDCl₃): δ 7.90 (d, 1H), 7.29 (t, 1H), 7.16 (d,1H), 6.76 (tt, 1H), 6.67-6.60 (m, 2H), 5.02-4.93 (m, 1H), 2.01 (br d,2H) 402

394 (M + NH₄) (400 MHz, CDCl₃): δ 8.08 (d, 1H), 7.31 (t, 1H), 7.10 (d,1H), 6.71 (tt, 1H), 6.61-6.54 (m, 2H), 5.52-5.48 (m, 1H), 4.05 (td, 1H),3.32 (ddd, 1H), 2.84-2.74 (m, 1H), 2.73-2.64 (m, 2H) 403

441, 443, 445 (M − H) (CDCl₃, 400 MHz) δ: 7.80 (d, 1H), 7.22 (d, 1H),7.02-7.00 (m, 1H), 6.87-6.86 (m, 1H), 6.72 (dt, 1 H), 5.38 (d, 1H) 404

475, 477, 479 (M + HCO2—) (CDCl₃, 400 MHz) δ: 8.00 (d, 1H), 7.46 (d,1H), 6.99-6.96 (m, 1H), 6.81-6.80 (m, 1H), 6.67 (dt, 1H), 2.08-2.04 (m,2H), 1.96-1.93 (m, 2H) 405

441, 443, 445 (M − H) (CDCl₃, 400 MHz): δ 7.80 (d, 1H), 7.22 (d, 1H),7.02-7.00 (m, 1H), 6.87-6.86 (m, 1H), 6.72 (dt, 1H), 5.38 (d, 1H) 406

479, 481, 483 (M + HCO₂ ⁻) (CDCl₃, 400 MHz): δ 7.73 (dd, 1H), 7.16 (d,1H), 6.98-6.95 (m, 1H), 6.83-6.82 (m, 1H), 6.66 (dt, 1H), 1.65 (s, 3H),1.40 (s, 3H) 407

352, 354 (M − H) (CDCl₃, 400 MHz): δ 7.89 (d, 1H), 7.13 (d, 1 H),7.09-7.06 (m, 1H), 6.96-6.94 (m, 1H), 6.80 (dt, 1 H), 5.78-5.74 (m, 1H),3.91 (dd, 1H), 3.65 (dd, 1H), 3.41 (d, 1H) 408

380, 382 (M − H) (CDCl₃, 400 MHz): δ 7.91 (d, 1H), 7.10-7.06 (m, 2H),6.97-6.96 (m, 1H), 6.81 (dt, 1H), 1.55 (s, 3H), 1.49 (s, 3H) 409

397 (M + NH₄) (400 MHz, CDCl₃): δ 7.85 (d, 1H), 7.27-7.23 (m, 2H), 7.15(s, 1H), 7.04 (d, 1H), 5.46 (dd, 1H), 3.69 (s, 1H), 3.13 (d, 1H) 410

410 (M − H) (400 MHz, CDCl₃): δ 7.85 (d, 1H), 7.28-7.25 (m, 1H), 7.23(d, 1H), 7.16- 7.14 (m, 1H), 7.05 (dt, 1H), 5.43 (d, 1H), 5.19 (d, 2H),3.27 (s, 1H) 411

394 (M + NH₄) (400 MHz, CDCl₃): δ 7.92 (d, 1H), 7.46-7.10 (m, 7H), 5.61(d, 1H), 5.26 (s, 2H), 3.00 (s, 1H) 412

407, 409 (M + H) (400 MHz, CDCl₃): δ 7.89-7.81 (m, 1H), 7.31-7.26 (m,1H), 6.78 (t, 1H), 6.65 (d, 2H), 5.59-5.51 (m, 1H), 4.60- 4.38 (br s,1H) 413

(400 MHz, CDCl₃): δ 7.72 (d, 1H), 7.21 (d, 1H), 6.71-6.64 (m, 1H), 6.57-6.52 (m, 2H), 5.60 (t, 1H), 3.83-3.78 (m, 1H), 3.71-3.67 (m, 1H), 2.88(d, 1H) 414

(400 MHz, CDCl₃): δ 7.89 (d, 1H), 7.15 (d, 1H), 6.83-6.76 (m, 1H), 6.71-6.67 (m, 2H), 5.78-5.74 (m, 1H), 3.94-3.89 (m, 1H), 3.67-3.63 (m, 1H),3.27-3.24 (m, 1H) 415

(400 MHz, CDCl₃): δ 7.77 (d, 1H), 7.25 (d, 1H), 7.22-7.19 (m, 1H), 7.06-7.05 (m, 1H), 6.99-6.96 (m, 1H), 5.62-5.59 (m, 1H), 3.85-3.80 (m, 1H),3.71-3.69 (m, 1H), 2.90 (d, 1H) 416

(400 MHz, CDCl₃): δ 7.94 (d, 1H), 7.35-7.31 (m, 1H), 7.24-7.22 (m, 1H),7.16 (d, 1H), 7.16-7.12 (m, 1H), 5.80- 5.73 (m, 1H), 3.96-3.90 (m, 1H),3.68-3.63 (m, 1H), 3.48-3.46 (m, 1H) 417

(400 MHz, CDCl₃): δ 8.24 (d, 1H), 7.89-7.86 (dd, 1H), 7.11 (d, 1H),6.70-6.55 (m, 1H), 6.59-6.51 (m, 2H), 3.10 (s, 2H) 418

(400 MHz, CDCl₃): δ 7.72 (d, 1H), 7.20 (d, 1H), 6.70-6.65 (m, 1H), 6.58-6.52 (m, 2H), 5.62-5.58 (m, 1H), 3.83-3.78 (m, 1H), 3.71-3.67 (m, 1H),2.93 (d, 1H) 419

(400 MHz, CDCl₃): δ 7.66 (d, 1H), 7.40-7.35 (m, 1H), 7.10 (d, 1H), 6.97-6.93 (m, 1H), 6.83-6.76 (m, 2H), 5.62-5.59 (m, 1H), 3.82-3.77 (m, 1H),3.70-3.66 (m, 1H), 2.96 (d, 1H) 420

(400 MHz, CDCl₃ + CD₃OD): δ 8.69 (d, 1H), 8.61-8.60 (d, 1H), 7.79 (d,1H), 7.55-7.54 (m, 1H), 7.29 (d, 1H), 5.57 (d, 1H), 3.84-3.79 (m, 1H),3.70- 3.66 (m, 1H) 421

(400 MHz, CDCl₃): δ 8.50 (s, 1H), 8.33 (s, 1H), 7.81 (d, 1H), 7.41-7.40(m, 1H), 7.19 (d, 1H), 5.41-5.37 (m, 1H), 3.45-3.40 (m, 1H) 422

(400 MHz, CDCl₃): δ 7.83 (d, 1H), 7.48-7.42 (m, 1H), 7.08-7.03 (m, 1H),7.06 (d, 1H), 6.94-6.87 (m, 2H), 5.78- 5.73 (m, 1H), 3.93-3.88 (m, 1H),3.66-3.62 (m, 1H), 3.56 (d, 1H) 423

(400 MHz, CDCl₃): δ 8.43 (d, 1H), 8.29 (d, 1H), 7.82 (d, 1H), 7.21 (d,1H), 7.18-7.14 (dt, 1H), 5.41-5.37 (m, 1H), 3.29-3.28 (m, 1H) 424

(400 MHz, CDCl₃): δ 8.76 (s, 1H), 8.64 (d, 1H), 7.87 (d, 1H), 7.59-7.58(m, 1H), 7.27 (d, 1H), 5.42-5.36 (m, 1H), 3.36-3.32 (m, 1H) 425

407 (M + HCOO—H) (400 MHz, CDCl₃): δ 7.81 (d, 1H), 7.28 (d, 1H),7.24-7.21 (m, 1H), 6.76- 6.70 (1H), 6.65-6.95 (m, 2H), 2.80 (m, 1H),1.75 (m, 3H) 426

437, 439 (M − H) (400 MHz, CDCl₃) δ 7.75 (d, 1H), 7.17 (d, 1H),6.56-6.51 (m, 1H), 6.42- 6.37 (m, 2H), 5.42-5.35 (m, 1H), 3.80 (s, 3H),2.99-2.95 (m, 1H) 427

384 (M − H) (400 MHz, CDCl₃) δ 7.92 (d, 1H), 7.17 (d, 1H), 6.64-6.60 (m,1H), 6.50- 6.45 (m, 2H), 6.57-6.51 (m, 1H), 3.82 (s, 3H), 3.80-3.74 (m,1H) 428

(400 MHz, CDCl₃): δ 8.38-8.36 (m, 1H), 8.14 (d, 1H), 7.72 (d, 1H), 7.24-7.22 (m, 1H), 7.01 (d, 1H), 5.41-5.37 (m, 1H), 4.14-4.08 (m, 1H), 2.40(s, 3H) 429

(400 MHz, CDCl₃): δ 8.26-8.23 (m, 1H), 7.95-7.93 (m, 1H), 7.73 (d, 1H),7.06 (d, 1H), 6.95-6.92 (m, 1H), 5.42- 5.35 (m, 1H), 4.03-3.97 (m, 1H),3.88 (s, 3H) 430

(400 MHz, CDCl₃): δ 7.80 (d, 1H), 7.59-7.54 (m, 2H), 7.37-7.35 (m, 1H),7.32-7.27 (m, 1H), 7.15 (d, 1H), 5.42- 5.37 (m, 1H), 3.11 (d, 1H) 431

388, 390 (M − H) (400 MHz, CDCl₃): δ 7.82 (d, 1H), 7.32 (d, 1H),7.25-7.23 (m, 1H), 7.12- 7.10 (m, 1H), 7.03-7.00 (dt, 1H), 5.44 (d, 1H),3.39-3.25 (m, 1H) 432

(400 MHz, CDCl₃): δ 8.50 (d, 1H), 8.33 (d, 1H), 7.78 (d, 1H), 7.41-7.40(m, 1H), 7.22 (d, 1H), 5.47-5.42 (m, 1H), 3.45 (d, 1H) 433

430 (M + HCOO—H) (400 MHz, CDCl₃): δ 7.62 (d, 1H), 7.32 (d, 1H),7.21-7.18 (m, 1H), 7.08- 7.06 (m, 1H), 6.97-6.93 (m, 1H), 5.47-5.43 (m,1H), 4.02 (s, 3H), 3.10- 3.08 (m, 1H) 434

418 (M + HCOO—H) (400 MHz, CDCl₃): δ 7.72 (d, 1H), 7.44-7.40 (m, 1H),7.25-7.22 (m, 1H), 7.13-7.10 (m, 1H), 7.05-7.01 (m, 1H), 5.51 (d, 1H)435

(400 MHz, CDCl₃): δ 7.92 (d, 1H), 7.62-7.57 (m, 2H), 7.42-7.40 (m, 1H),7.35-7.31 (m, 1H), 7.28 (t, J = 53 Hz, 1H), 7.11 (d, 1H), 5.69-5.65 (m,1H), 3.33-3.32 (m, 1H) 436

(400 MHz, CDCl₃ + CD₃OD): δ 7.80 (d, 1H), 7.23-7.15 (m, 1H), 7.20 (t,1H), 7.02 (d, 1H), 6.94-6.88 (m, 1H), 6.80-6.75 (m, 1H), 5.50 (d, 1H)437

(400 MHz, CDCl₃ + CD₃OD): δ 7.83 (d, 1H), 7.33-7.23 (m, 3H), 7.18 (t,1H), 7.01 (d, 1H), 5.51-5.48 (m, 1H) 438

(400 MHz, CDCl₃): δ 7.93 (d, 1H), 7.24 (d, 1H), 7.21 (t, 1H), 5.63-5.58(m, 1H), 4.23 (q, 2H), 2.94-2.89 (m, 1H), 1.51 (t, 3H) 439

(400 MHz, CDCl₃): δ 7.92 (d, 1H), 7.26 (t, 1H), 7.21 (d, 1H), 5.63-5.59(m, 1H), 4.01-3.98 (m, 2H), 2.95-2.92 (m, 1H), 1.37-1.27 (m, 1H),0.74-0.70 (m, 2H), 0.42-0.38 (m, 2H) 440

(400 MHz, CDCl₃): δ 7.85 (d, 1H), 7.78 (d, 1H), 7.50 (d, 1H), 7.47 (t,1H), 7.42 (t, 1H), 7.17 (dd, 1H), 7.05 (dd, 1H), 6.96 (d, 1H), 5.73-5.67(m, 1H), 3.20-3.13 (br s, 1H) 441

(400 MHz, CDCl₃): δ 8.17 (s, 1H), 7.83 (d, 1H), 7.72 (d, 1H), 7.42 (t,1H), 7.22 (t, 1H), 7.12-7.07 (m, 2H), 5.69 (d, 1H) 442

418 (M − H) (400 MHz, CDCl₃): δ 7.95-7.90 (d, 1H), 7.65 (t, 1H),7.25-7.22 (m, 1H), 7.22-7.18 (d, 1H), 7.13-7.11 (m, 1H), 7.08-7.04 (m,1H), 3.73 (brd, s, 1H), 1.91-1.88 (m, 3H) 443

372 (M − H) (400 MHz, CDCl₃): δ 7.97 (d, 1H), 7.20 (d, 1H), 6.87-6.81(m, 1H), 6.76- 6.69 (m, 2H), 5.55 (dd, 1H) 444

439, 441 (M − H) (400 MHz, CDCl₃): δ 7.78 (d, 1H), 7.16 (d, 1H),6.75-6.69 (m, 1H), 6.62- 6.54 (m, 2H), 3.04-2.98 (m, 1H), 1.95 (d, 3H)445

472 (M − H) (400 MHz, CDCl₃): δ 8.03 (d, 1H), 7.57 (t, 1H), 7.37 (d,1H), 7.29-7.25 (m, 1H), 7.17-7.14 (m, 1H), 7.08 (dt, 1H), 5.40-5.10 (m,1H) 446

386 (M − H) (400 MHz, CDCl₃): δ 7.93 (d, 1H), 7.13 (d, 1H), 6.83 (tt,1H), 6.75-6.68 (m, 2H), 3.54 (s, 1H), 1.97 (d, 3H) 447

457, 459 (M − H) (400 MHz, CDCl₃): δ 7.79 (d, 1H), 7.27 (d, 1H), 6.72(tt, 1H), 6.62-6.55 (m, 2H), 4.71 (s, 1H), 4.16 (d, 1H), 3.83 (d, 1H)448

404 (M − H) (400 MHz, CDCl₃): δ 7.94 (d, 1H), 7.26 (d, 1H), 6.83 (tt,1H), 6.76-6.69 (m, 2H), 4.26 (br s, 1H), 4.18 (d, 1H), 3.81 (d, 1H) 449

464, 466 (M − H) (400 MHz, CDCl₃): δ 7.84 (d, 1H), 7.30 (d, 1H),7.27-7.24 (m, 1H), 7.12- 7.10 (m, 1H), 7.00 (dt, 1H), 4.63 (s, 1H), 4.18(d, 1H), 3.85 (d, 1H) 450

411 (M − H) (400 MHz, CDCl₃): δ 7.99 (d, 1H), 7.38-7.34 (m, 1H),7.29-7.24 (m, 2H), 7.20-7.16 (dt, 1H), 4.75 (br s, 1H), 4.20 (d, 1H),3.82 (d, 1H) 451

393 (M − H) (400 MHz, CDCl₃): δ 7.98 (d, 1H), 7.39-7.35 (m, 1H),7.27-7.25 (m, 1H), 7.17 (dt, 1H), 7.13 (d, 1H), 3.27 (d, 1H), 1.99 (d,3H) 452

401 (M + NH₄ ⁺) (400 MHz, CDCl₃): δ 7.89 (d, 1H), 7.27-7.24 (m, 1H),7.21 (t, 1H), 7.16- 7.11 (m, 2H), 7.06-7.02 (m, 1H), 5.34 (br s, 1H),3.66-3.58 (m, 1H), 3.13 (br s, 1H), 1.55 (d, 3H) 453

401 (M + NH₄ ⁺) (400 MHz, DMSO-d6): δ 7.99 (d, 1H), 7.75-7.71 (m, 1H),7.52-7.49 (m, 1H), 7.48-7.44 (m, 1H), 7.36 (d, 1H), 7.29 (t, 1H), 6.23(d, 1H), 5.52 (t, 1H), 3.76-3.68 (m, 1H), 1.35 (d, 3H) 454

347 (M + NH₄ ⁺) (400 MHz, CDCl₃): δ 7.84 (d, 1H), 7.31 (t, 1H), 7.09 (d,1H), 5.87-5.79 (m, 1H), 4.17-4.04 (m, 2H), 3.75-3.68 (m, 1H), 3.65-3.59(m, 1H), 2.91-2.84 (m, 1H), 1.54-1.48 (m, 2H), 1.22-1.14 (m, 2H) 455

410 (M + HCO₂ ⁻) (400 MHz, CDCl₃): δ 7.86 (d, 1H), 7.19 (t, 1H), 7.14(d, 1H), 5.47 (d, 1H), 4.15-4.03 (m, 2H), 3.15 (s, 1H), 1.47-1.39 (m,2H), 1.19-1.12 (m, 2H) 456

316 (M − H) (400 MHz, CDCl₃): δ 7.92 (d, 1H), 7.18 (d, 1H), 4.81 (m,1H), 3.18 (s, 1H), 1.92 (d, 3H), 1.48 (t, 6H) 457

393 (M − H) (400 MHz, CDCl₃): δ 7.98 (d, 1H), 7.39-7.35 (m, 1H),7.27-7.25 (m, 1H), 7.17 (dt, 1H), 7.13 (d, 1H), 3.27 (d, 1H), 1.99 (d,3H) 458

393 (M − H) (400 MHz, CDCl₃): δ 7.98 (d, 1H), 7.39-7.35 (m, 1H),7.27-7.25 (m, 1H), 7.17 (dt, 1H), 7.13 (d, 1H), 3.27 (d, 1H), 1.99 (d,3H) 459

410 (M + HCO2−) (400 MHz, (CD₃)₂CO): δ 8.23 (d, 1H), 7.74 (d, 1H), 7.57(d, 2H), 7.48- 7.36 (m, 3H), 6.54 (d, 1H), 5.54 (s, 2H), 1.86 (d, 3H)460

405 (M − H) (400 MHz, CDCl₃): δ 8.00 (d, 1H), 7.39-7.35 (m, 1H),7.26-7.24 (m, 1H), 7.18-7.13 (m, 2H), 6.12 (ddd, 1H), 5.86 (d, 1H), 5.73(d, 1H), 3.45 (d, 1H) 461

364 (M − H) (400 MHz, CDCl₃): δ 7.97 (d, 1H), 7.02 (d, 1H), 4.90-4.81(m, 1H), 3.29- 3.17 (m, 2H), 3.10 (d, 1H), 3.02-2.87 (m, 2H), 1.93 (d,3H) 462

486, 488, 490 (M + H) (400 MHz, (CD₃)₂SO): δ 8.15 (d, 1H), 7.48 (d, 1H),7.47 (ddd, 1H), 7.29- 7.27 (m, 1H), 7.18 (dt, 1H), 5.04 (d, 1H) 463

342 (M − H) (400 MHz, (CD₃)₂CO): δ 7.99 (d, 1H), 7.66-7.61 (m, 2H), 7.56(dt, 1H), 7.42 (d, 1H), 5.03 (br s, 1H), 3.95 (dd, 1H), 3.48 (dd, 1H),2.40 (br s, 2H) 464

380 (M + H) (400 MHz, (CD₃)₂CO): δ 8.20 (d, 1H), 7.62 (ddd, 1H),7.55-7.53 (m, 1H), 7.45 (dt, 1H), 7.39 (d, 1H), 5.14- 5.04 (m, 1H), 2.42(br d, 2H) 465

(M + H) 328 (400 MHz, CDCl₃): δ 8.04 (s, 1H), 5.46-5.26 (m, 2H),4.89-4.79 (m, 1H), 3.36-3.08 (m, 4H), 2.91-2.74 (m, 2H), 2.60 (dd, 1H)466

(M + H) 310 (400 MHz, CDCl₃): δ 7.98 (s, 1H), 5.59-5.54 (m, 1H),4.88-4.79 (m, 1H), 3.24-3.07 (m, 3H), 2.89 (dd, 1H), 2.89-2.74 (m, 2H),2.44-2.34 (m, 1H), 2.28-2.21 (m, 1H), 2.12-2.09 (m, 1H) 467

(M + H) 357 (400 MHz, CDCl₃): δ 8.33 (s, 1H), 7.22 (ddd, 1H), 7.10-7.08(m, 1H), 6.99 (dt, 1H), 5.54-5.46 (m, 1H), 5.46-5.28 (m, 1H), 3.26 (ddd,1H), 3.11 (ddd, 1H), 2.67 (dd, 1H) 468

(M + H) 367 (400 MHz, CDCl₃): δ 8.27 (s, 1H), 7.25 (ddd, 1H), 7.14 (m,1H), 7.03 (dt, 1H), 5.69 (dt, 1H), 5.53-5.36 (m, 1H), 4.24 (d, 1H), 3.34(s, 3H), 3.31- 3.24 (m, 1H), 3.09 (ddd, 1H) 469

326/328 (M + H) 470

362 (M + H) (400 MHz, CDCl₃): δ 7.49 (s, 1H), 6.80-6.71 (m, 3H), 6.19(t, 1H), 3.36 (t, 2H), 3.06 (t, 2H), 2.31-2.23 (m, 2H) 471

(M + H) 341 (400 MHz, CDCl₃): δ 8.65 (s, 1H), 7.53-7.48 (m, 2H), 7.40(ddd, 1H), 5.56-5.48 (m, 1H), 5.35 (ddt, 1H), 3.41 (ddd, 1H), 3.21 (ddd,1H), 2.65 (dd, 1H) 472

339 (M + H) 473

302 (M + H) 474

284 (M + H) 475

338 (M + H) 476

338 (M + H) 477

333 (M + H) 478

310 (M + H) 479

340 (M + H) 480

(M + HCO₂ ⁻) 445 (400 MHz, CDCl₃): δ 7.92 (d, 1H), 7.52 (d, 1H), 5.40(dd, 1H), 4.33-4.28 (m, 1H), 3.59 (ddd, 1H), 3.49 (t, 1H), 3.22-3.13 (m,2H), 1.97-1.82 (m, 4H), 1.75-1.58 (m, 3H), 1.46 (dd, 1H), 1.42-1.37 (m,1H) 481

(M − OH) 381 (400 MHz, CDCl₃): δ 7.93 (d, 1H), 7.50 (d, 1H), 5.91-5.88(m, 1H), 5.39 (dd, 1H), 4.49-4.43 (m, 1H), 3.64 (ddd, 1H), 3.39 (t, 1H),3.18 (dd, 1H), 2.49-2.39 (m, 1H), 2.22-2.11 (m, 1H), 2.09-1.92 (m, 2H),1.81-1.59 (m, 3H) 482

(M + HCO₂ ⁻) 445 (400 MHz, CDCl₃): δ 7.94 (d, 1H), 7.55 (d, 1H), 5.41(dd, 1H), 3.82-3.72 (m, 1H), 3.56 (ddd, 1H), 3.42 (t, 1H), 3.20 (dd,1H), 2.67 (tt, 1H), 2.17-2.08 (m, 2H), 2.01-1.94 (m, 1H), 1.82-1.75 (m,1H), 1.60-1.42 (m, 3H), 1.40-1.27 (m, 2H) 483

(M + HCO₂ ⁻) 445 (400 MHz, CDCl₃): δ 7.94 (d, 1H), 7.55 (d, 1H), 5.41(dd, 1H), 3.82-3.72 (m, 1H), 3.58 (ddd, 1H), 3.40 (t, 1H), 3.18 (d, 1H),2.67 (tt, 1H), 2.16-2.06 (m, 2H), 2.02-1.95 (m, 1H), 1.86-1.78 (m, 1H),1.58-1.28 (m, 5H) 484

366 (M + H) 485

(M − H) 411/413 486

(M − H) 420 487

(M − H) 377 488

(M + H) 420 489

(M − H) 435 490

(M + H) 388 491

(M + Na) 449 492

(M + Na) 422 493

453/455 (M + HCO₂ ⁻) 494

346 (M − H) 495

309 (M − H) 496

345 (M − H) 497

320 (M − H) 498

345 (M − H) 499

337 (M − H) 500

373 (M − H) 501

356 (M − H) 502

381 (M − H) 503

391 (M − H) 504

408 (M − H) 505

399 (M − H) 506

392 (M − H) 507

¹HNMR (300 MHz, CDCl₃): δ 7.83 (d, 1H), 7.30 (d, 1H), 6.77 (m, 2H), 6.67(m, 1H), 6.10 (s, 1H), 5.36 (m, 1H), 3.45 (m, 1H), 3.27 (m, 2H). 508

361 (M − H) 509

397 (M − H) 510

355 (M − H) 511

391 (M − H) 512

¹HNMR (300 MHz, CDCl₃): δ 8.76 (d, 1H), 6.97 (d, 1H), 5.62 (m, 1H), 3.29(m, 1H), 3.09 (m, 1H), 3.05 (m, 1H), 2.36 (m, 2H), 1.98 (m, 1H), 1.17(m, 2H), 0.85 (m, 2H). 513

341 (M − H) 514

294 (M − H) 515

307 (M − H) 516

329 (M − H) 517

343 (M − H) 518

322 (M − H) 519

214 (M + H) 520

345 (M − H) 521

375 (M − H) 522

323 (M − H) 523

322 (M − H) 524

323 (M − H) 525

323 (M − H) 526

360 (M + H) 527

360 (M + H) 528

(400 MHz, CDCl₃): δ 7.79 (d, 1H), 7.29 (d, 1H), 7.07-7.00 (m, 2H), 6.78(d, 1H), 6.33 (t, 1H), 5.40 (d, 1H), 4.66 (d, 2H), 4.64-4.58 (m, 1H),3.38- 3.24 (m, 2H), 3.14 (t, 1H) 529

(400 MHz, CDCl₃): δ 7.79 (d, 1H), 7.40 (s, 1H), 6.80 (d, 1H), 6.46-6.20(m, 3H), 5.39 (d, 1H), 4.64 (t, 1H), 4.47 (d, 2H), 3.46-3.24 (m, 2H),3.14 (t, 1H) 530

(400 MHz, CDCl₃): δ 7.79 (d, 1H), 7.28 (d, 1H), 7.01-6.94 (m, 3H), 6.35(t, 1H), 5.38 (d, 1H), 4.61 (m, 2H), 3.70-3.58 (m, 1H), 3.52-3.44 (m,1H), 3.23 (br s, 1H), 3.00 (s, 3H) 531

[M − H]⁻ 392 532

[M + H]⁺ 343 533

(400 MHz, CDCl₃): δ 8.00 (dd, 1H), 7.84 (dd, 1H), 5.41 (dd, 1H), 5.00-4.93 (m, 1H), 3.83 (ddd, 1H), 3.72- 3.43 (m, 4H) 534

[M − H]⁻ 382 535

[M − H]⁻ 382 536

[M − H]⁻ 382 537

[M − H]⁻ 331 538

[M − H]⁻ 406 539

[M + H]⁺ 395 540

[M + H]⁺ 428/430 541

[M + H]⁺ 425 542

[M + NH₄]⁺ 444 543

[M − OH]⁺ 409 544

[M + Cl]⁻ 463/465 545

[M + Cl]⁻ 463/465 546

[M + NH₄]⁺ 444 547

[M + Cl]⁻ 463/465 548

[M + NH4]⁺ 446 549

[M + NH4]⁺ 446 550

[M + NH4]⁺ 446 551

[M − OH]⁺ 409 552

[M + NH4]⁺ 446 553

[M + H]⁺ 366 554

[M + NH₄]⁺ 310 555

[M + NH₄]⁺ 346 556

[M + NH₄]⁺ 346 557

[M + Na]⁺ 454 558

[M + H]⁺ 380 559

[M + H]⁺ 383 560

[M + H]⁺ 369 561

[M + H]⁺ 369 562

[M + H]⁺ 397 563

[M + Na]⁺ 436/438 564

[M − H]⁻ 393 565

[M + Na]⁺ 436/438 566

[M + H]⁺ 383 567

[M + H]⁺ 434 568

[M + H]⁺ 419 569

[M − H]⁻ 402 570

[M + H]⁺ 405 571

(M − H) 381 572

(M − H) 407 573

(M − H) 405 574

373 (M + H) 575

373 (M + H) 576

(M + H) 391 577

(M + NH4) 372 578

(M + NH4) 390 579

[M − H + formate] 363 580

[M + H] 332 581

(400 MHz, CDCl₃): δ 8.33 (d, 1H), 7.88-7.84 (m, 1H), 7.04 (d, 1H), 5.82(dd, 1H), 4.67 (dt, 2H), 4.33-4.24 (m, 2H), 2.32-2.20 (m, 2H) 582

(400 MHz, CDCl₃): δ 7.80-7.77 (m, 1H), 7.05 (d, 1H), 5.74 (dd, 1H),5.25-5.20 (m, 1H), 4.66 (dt, 2H), 4.33-4.22 (m, 2H), 2.52 (d, 1H), 2.32-2.19 (m, 2H) 583

(300 MHz, CDCl₃): δ 7.87 (d, 1H), 6.95 (d, 1H), 5.58 (d, 1H), 4.08 (m,2H), 2.06-3.17 (m, 2H), 2.84-2.94 (m, 1H), 2.37 (m, 1H), 2.27 (m, 1H),1.86 (m, 2H), 1.07 (t, 3H) 584

359 (M − H) 585

337 (M + H) 586

345 (M − H + HCOOH) 587

389 (M − H + HCOOH) 588

(300 MHz, CDCl₃): δ 7.90 (d, 1H), 6.97 (d, 1H), 5.58 (m, 1H), 3.97 (s,3H), 3.17 (m, 1H), 3.12 (m, 1H), 2.87 (m, 1H), 2.37 (m, 1H), 2.27 (m,1H). 589

(300 MHz, CDCl₃): δ 7.86 (d, 1H), 6.94 (d, 1H), 5.58 (m, 1H), 4.20 (m,2H), 3.17 (m, 1H), 3.09 (m, 1H), 2.89 (m, 1H), 2.35 (m, 1H), 2.26 (m,1H), 1.47 (t, 3H). 590

331 (M − H) 591

345 (M − H) 592

(300 MHz, CDCl₃): δ 7.85 (d, 1H), 6.94 (d, 1H), 5.57 (m, 1H), 4.72 (m,1H), 3.18 (m, 1H), 3.11 (m, 1H), 2.86 (m, 1H), 2.35 (m, 1H), 2.25 (m,1H), 1.41 (d, 6H). 593

359 (M − H) 594

389 (M − H) 595

375 (M − H) 596

(300 MHz, CD₃OD): δ 7.89 (d, 1H), 7.28 (d, 1H), 5.44 (m, 1H), 5.27 (m,1H), 5.14 (d, 1H), 3.77-3.96 (m 4H), 2.93 (m, 1H), 2.76 (m, 1H), 2.29(m, 1H), 2.15 (m, 1H), 1.97 (m, 2H). 597

(300 MHz, CDCl₃): δ 7.85 (d, 1H), 6.92 (d, 1H), 5.57 (m, 1H), 3.96 (m,2H), 3.17 (m, 1H), 3.08 (m, 1H), 2.94 (m, 1H), 2.36 (m, 1H), 2.26 (m,1H), 1.28 (m, 1H), 0.68 (m, 2H), 0.39 (m, 2H). 598

371 (M − H) 599

(300 MHz, CDCl₃): δ 7.87 (d, 1H), 6.99 (d, 1H), 5.57 (m, 1H), 4.27 (m,2H), 3.80 (m, 2H), 3.45 (s, 3H), 3.15 (m, 1H), 3.11 (m, 1H), 2.93 (m,1H), 2.36 (m, 1H), 2.27 (m, 1H) 600

372 (M + NH₄) 601

(300 MHz, CDCl₃): δ 7.82 (d, 1H), 6.79 (d, 1H), 5.57 (m, 1H), 4.78 (m,1H), 3.17 (m, 1H), 3.09 (m, 1H), 2.89 (m, 1H), 2.50 (m, 2H), 2.20-2.28(m, 4H), 1.77 (m, 1H), 1.55 (m, 1H). 602

371 (M − H) 603

(300 MHz, CDCl₃): δ 7.85 (d, 1H), 6.95 (d, 1H), 5.56 (m, 1H), 4.91 (m,1H), 3.18 (m, 1H), 3.10 (m, 1H), 2.87 (m, 1H), 2.32 (m, 1H), 2.25 (m,1H), 1.56-1.99 (m, 8H). 604

387 (M − H) 605

385 (M − H) 606

(300 MHz, CDCl₃): δ 7.92 (d, 1H), 7.04 (d, 1H), 5.36 (d, 1H), 4.60 (m,1H), 3.46 (m, 1H), 3.39 (m, 1H), 3.14 (m, 2H), 2.80 (m, 2H), 2.05 (m,2H), 1.76 (m, 2H), 1.55 (m, 4H) 607

402 (M + H) 608

389 (M − H) 609

(300 MHz, CDCl₃): δ 7.90 (d, 1H), 6.94 (d, 1H), 5.60 (m, 1H), 4.04 (m,2H), 3.20 (m, 1H), 3.15 (m, 1H), 2.98 (m, 1H), 2.40 (m, 1H), 2.31 (m,1H), 1.54 (s, 3H), 1.53 (s, 3H) 610

398 (M − H) 611

(300 MHz, CDCl₃): δ 7.90 (d, 1H), 6.94 (d, 1H), 5.60 (m, 1H), 4.04 (m,2H), 3.20 (m, 1H), 3.15 (m, 1H), 2.98 (m, 1H), 2.40 (m, 1H), 2.31 (m,1H), 1.54 (s, 3H), 1.53 (s, 3H) 612

377 (M + H) 613

371 (M − H + HCOOH) 614

363 (M + H) 615

425 (M − H + HCOOH) 616

382 (M + NH₄ ⁺) 617

399 (M − H) 618

415 (M − H) 619

406 (M − H + HCOOH) 620

398 (M + H) 621

411 (M − H + HCOOH) 622

401 (M − H) 623

415 (M − H) 624

425 (M − H + HCOOH) 625

372 (M + NH₄) 626

389 (M − H) 627

444 (M + H) 628

480 (M + H) 629

430 (M + H) 630

466 (M + H) 631

439 (M + H) 632

404 (M + H) 633

377 (M + H) 634

416 (M + H) 635

362 (M + H) 636

404 (M + H) 637

440 (M + H) 638

390 (M + H) 639

389 (M − H) 640

374 (M − H) 641

390 (M + H) 642

(300 MHz, CDCl₃): δ 7.93 (d, 1H), 6.94 (d, 1H), 5.35 (m, 1H), 4.96 (m,1H), 3.38-3.47 (m, 3H), 2.82-2.92 (m 3H), 2.52 (m, 1H), 2.43 (m 1H),2.42 (s, 3H), 2.04 (m 1H) 643

388 (M + H) 644

433 (M − H + HCOOH) 645

402 (M − H) 646

389 (M − H) 647

418 (M + H) 648

(300 MHz, CDCl₃): δ 7.92 (d, 1H), 6.87 (d, 1H), 5.36 (m, 1H), 4.93 (m,1H), 3.80 (s, 3H), 3.44-3.60 (m, 2H), 3.22 (m, 1H), 1.71 (d, 3H) 649

388 (M − H) 650

376 (M + H) 651

(400 MHz, CDCl₃): δ 7.31 (m, 1H), 7.17 (d, 1H), 6.82 (d, 1H), 6.09 (t,1H), 5.15 (m, 1H), 4.20 (m, 2H), 3.45 (m, 1H), 3.27 (m, 1H), 2.27 (m,1H) 652

313 (M − H + HCOOH) 653

(400 MHz, CDCl₃): δ 7.56 (d, 1H), 7.17 (d, 1H), 5.13 (m, 1H), 3.56 (m,2H), 2.50 (s, 1H) 654

359 (M − H) 655

359 (M − H) 656

371 (M − H) 657

371 (M − H) 658

442 (M + HCO₂ ⁻) 659

389 (M − H) 660

389 (M − H) 661

424 (M + HCO₂ ⁻) 662

265 (M − OH) 663

(M + NH4) 426 664

(M − H) 421 665

(M + H) 410 666

(400 MHz, CDCl₃): δ 8.15 (s, 1H), 6.12 (t, 1H), 4.96-4.87 (1H), 4.22-4.09 (2H), 3.62-3.50 (2H), 2.49 (1H) 667

(M − OH) 467 668

(M + NH4) 330 669

(M + H) 350 670

(400 MHz, CDCl₃): δ 7.63 (d, 1H), 7.25 (d, 1H), 6.62 (t, 1H), 5.30-5.25(m, 1H), 3.58-3.57 (m, 2H), 2.54-2.51 (m, 1H) 671

(400 MHz, CDCl₃): δ 7.59 (d, 1H), 7.18 (d, 1H), 6.60 (t, 1H), 5.40-5.23(m, 2H), 3.40-3.12 (m, 2H), 2.55-2.51 (m, 1H) 672

(M + H) 358 673

[M − OH]⁺ 229 674

[M − H + HCOOH]⁻ 284 675

[M − H + HCOOH]⁻ 445 676

[M + NH4]⁺ 360 677

[M + NH4]⁺ 382 678

[M + NH4]⁺ 346 679

(400 MHz, CDCl₃): δ 7.62 (d, 1H), 6.63 (d, 1H), 4.94 (dd, 1H), 3.81 (s,3H), 3.46-3.38 (m, 2H), 2.43-2.40 (m, 1H) 680

[M + Na]⁺ 289 681

(400 MHz, CDCl₃): δ 7.64 (d, 1H), 6.61 (d, 1H), 6.07 (tt, 1H), 4.94 (d,1H), 4.19 (td, 2H), 3.50-3.42 (m, 2H), 2.41 (brs, 1H) 682

(400 MHz, CDCl₃): δ 7.59 (d, 1H), 6.61 (d, 1H), 4.98-4.91 (m, 1H), 3.92(td, 2H), 3.48-3.39 (m, 2H), 2.40 (brs, 1H), 1.84-1.76 (m, 2H), 1.02 (t,3H) 683

(400 MHz, CDCl₃): δ 7.58 (d, 1H), 6.57 (d, 1H), 5.26 (dq, 1H), 5.06-5.01(m, 1H), 3.91 (t, 2H), 3.21 (dd, 2H), 2.42-2.38 (m, 1H), 1.84-1.74 (m,2H), 1.03 (t, 3H) 684

[M + H]⁺ 317 685

[M + NH4]⁻ 251 686

[M − H + HCOOH]⁻ 363 687

(400 MHz, CDCl₃): δ 7.42 (d, 1H), 6.73 (d, 1H), 6.07 (tt, 1H), 5.08 (d,1H), 4.23-4.17 (m, 2H), 3.54-3.35 (m, 2H), 2.49 (brs, 1H) 688

[M + H]⁺ 276 689

(400 MHz, CDCl₃): δ 7.78 (d, 1H), 7.09 (d, 1H), 6.17 (tt, 1H), 5.79 (dd,1H), 5.24-5.18 (m, 1H), 4.42-4.31 (m, 2H), 2.47-2.44 (m, 1H) 690

(400 MHz, CDCl₃): δ 7.74 (d, 1H), 7.05 (d, 1H), 6.16 (tt, 1H), 6.02 (dd,1H), 5.54-5.48 (m, 1H), 4.35 (td, 2H), 2.68 (brd, 1H) 691

[M − H]⁻ 290 692

[M + H]⁺ 310 693

[M + H]⁺ 286 694

[M − H + HCOOH]⁻ 343 695

[M + H]⁺ 317 696

(400 MHz, CDCl₃): δ 7.56 (d, 1H), 6.84 (d, 1H), 6.11 (tt, 1H), 5.38-5.22(m, 2H), 4.26 (td, 2H), 3.33-3.07 (m, 2H), 2.52-2.48 (m, 1H) 697

(400 MHz, CDCl₃): δ 7.56 (d, 1H), 6.87 (d, 1H), 6.12 (tt, 1H), 5.49-5.22(m, 2H), 4.31-4.23 (m, 2H), 3.48-3.06 (m, 2H), 2.48 (brs, 1H) 698

(400 MHz, CDCl₃): δ 7.62 (d, 1H), 7.61 (s, 1H), 6.81 (d, 1H), 6.13 (tt,1H), 4.28 (td, 2H), 3.09-2.97 (m, 4H) 699

[M − H + HCOOH]⁻ 381 700

[M − H]⁻ 344 701

[M − H + HCOOH]⁻ 377 702

[M + H]⁺ 328 703

[M + H]⁺ 310 704

[M + Na]⁺ 320 705

[M + H]⁺ 312 706

363 (M − H)⁻ 707

472 (M + HCO₂)⁻ 708

491 (M + HCO₂)⁻ 709

373 (M − H)⁻ 710

373 (M − H)⁻ 711

355 (M − H)⁻ 712

377 (M − H)⁻ 713

387 (M − H)⁻ 714

401 (M − H)⁻ 715

401 (M − H)⁻ 716

385 (M − H)⁻ 717

387 (M − H)⁻ 718

413 (M − H)⁻ 719

416 (M + H)⁺ 720

416 (M + H)⁺ 721

495 (M + HCO₂)⁻ 722

388 (M + H − CO₂—C₄H₈)⁺ 723

451 (M − H)⁻ 724

388 (M + H)⁺ 725

430 (M + H)⁺ 726

510 (M + HCO₂)⁻ 727

407 (M − H)⁻ 728

407 (M − H)⁻ 729

439 (M + H)⁺ 730

401 (M − H)⁻ 731

401 (M − H)⁻ 732

439 (M + H)⁺ 733

436 (M + H)⁺ 734

355 (M + H)⁺ 735

353 (M + H)⁺ 736

355 (M + H)⁺ 737

355 (M + H)⁺ 738

353 (M + H)⁺ 739

402 (M + Na)⁺ 740

355 (M + H)⁺ 741

380 (M + H)⁺ 742

390 (M + NH₄)⁺ 743

372 (M + NH₄)⁺ 744

[M + NH₄]⁺ 420 745

[M + NH₄]⁺ 384 746

[M + NH₄]⁺ 404 747

[M + NH₄]⁺ 422 748

[M + NH₄]⁺ 429 749

[M + NH₄]⁺ 445 750

[M + NH₄]⁺ 408 751

[M + NH₄]⁺ 382 752

[M + NH₄]⁺ 381 753

[M + NH₄]⁺ 354 754

[M + NH₄]⁺ 379 755

[M + NH₄]⁺ 364 756

[M + NH₄]⁺ 390 757

[M + NH₄]⁺ 407 (400 MHz, CDCl₃): δ 8.09 (d, 1H), 7.07 (d, 1H), 5.82-5.66(m, 1H), 5.61- 5.57 (m, 1H), 4.73 (m, 1H), 3.41 (d, 1H), 3.20 (s, 3H),2.87 (m, 1H), 2.17- 1.80 (m, 8H) 758

[M + NH₄]⁺ 407 (400 MHz, CDCl₃): δ 8.07 (dd, 1H), 7.04 (d, 1H),5.85-5.70 (m, 1H), 5.62- 5.58 (m, 1H), 4.65 (m, 1H), 3.25 (d, 1H), 3.20(s, 3H), 2.76-2.74 (m, 1H), 2.12-1.55 (m, 8H) 759

[M + NH₄]⁺ 404 760

[M + NH₄]⁺ 379 (400 MHz, CDCl₃): δ 8.10 (dd, 1H), 6.86 (d, 1H),5.82-5.66 (m, 1H), 5.61- 5.58 (m, 1H), 5.17-5.14 (m, 1H), 3.32-3.29 (m,1H), 3.24 (d, 1H), 3.20 (s, 3H), 2.99-2.93 (m, 2H), 2.76-2.70 (m, 2H)761

[M + NH₄]⁺ 379 (400 MHz, CDCl₃): δ 8.08 (dd, 1H), 6.84 (d, 1H),5.85-5.69 (m, 1H), 5.62- 5.58 (m, 1H), 4.86-4.79 (m, 1H), 3.24 (d, 1H),3.20 (s, 3H), 3.07-3.00 (m, 2H), 2.96-2.87 (m, 1H), 2.76-2.66 (m, 2H)762

[M + NH₄]⁺ 378 763

[M + NH₄]⁺ 397 (400 MHz, CDCl₃): δ 8.06 (dd, 1H), 6.90 (d, 1H),5.85-5.70 (m, 1H), 5.59- 5.56 (m, 1H), 5.39-5.32 (m, 2H), 4.79-4.76 (m,1H), 3.24 (d, 1H), 3.20 (s, 3H), 2.83-2.74 (m, 3H), 2.63-2.55 (m, 2H)764

[M + NH₄]⁺ 379 (400 MHz, CDCl₃): δ 8.10 (dd, 1H), 7.09 (d, 1H),5.91-5.76 (m, 1H), 5.58- 5.55 (m, 1H), 4.48-4.44 (m, 1H), 4.14-4.10 (m,1H), 3.41 (d, 1H), 3.20 (s, 3H), 1.94-1.91 (m, 1H), 1.81-1.76 (m, 1H),1.46-1.41 (m, 1H), 1.20-1.18 (m, 1H) 765

[M + NH₄]⁺ 379 (400 MHz, CDCl₃): δ 8.12 (dd, 1H), 7.10 (d, 1H),5.91-5.75 (m, 1H), 5.62- 5.58 (m, 1H), 4.39-4.29 (m, 2H), 3.26 (d, 1H),3.21 (s, 3H), 1.90-1.86 (m, 1H), 1.79-1.76 (m, 1H), 1.45-1.39 (m, 1H),1.26-1.22 (m, 1H) 766

[M + NH₄]⁺ 393 (400 MHz, CDCl₃): δ 8.10 (dd, 1H), 6.88 (d, 1H),5.82-5.66 (m, 1H), 5.61- 5.58 (m, 1H), 5.08-5.02 (m, 1H), 3.29 (d, 1H),3.20 (s, 3H), 3.19-3.14 (m, 2H), 2.42-2.32 (m, 2H), 1.65 (s, 3H) 767

[M + NH₄]⁺ 393 (400 MHz, CDCl₃): δ 8.05 (d, 1H), 6.81 (d, 1H), 5.87-5.71(m, 1H), 5.61- 5.58 (m, 1H), 4.97-4.91 (m, 1H), 3.25 (d, 1H), 3.20 (s,3H), 2.92-2.76 (m, 2H), 2.74-2.70 (m, 2H), 1.65 (s, 3H) 768

[M + NH₄]⁺ 390 (400 MHz, CDCl₃): δ 8.11 (dd, 1H), 7.07 (d, 1H),5.85-5.69 (m, 1H), 5.62- 5.58 (m, 1H), 4.36-4.32 (m, 1H), 4.21-4.17 (m,1H), 3.27 (d, 1H), 3.20 (s, 3H), 2.18-2.10 (m, 1H), 1.72-1.67 (m, 2H),1.41-1.33 (m, 1H) 769

[M + NH₄]⁺ 390 (400 MHz, CDCl₃): δ 8.11 (dd, 1H), 7.06 (d, 1H),5.87-5.72 (m, 1H), 5.61- 5.57 (m, 1H), 4.25-4.23 (m, 2H), 3.27 (d, 1H),3.20 (s, 3H), 2.22-2.10 (m, 1H), 1.73-1.70 (m, 2H), 1.39-1.36 (m, 1H)770

[M + NH₄]⁺ 379 (400 MHz, CDCl₃): δ 8.10 (dd, 1H), 7.02 (d, 1H),5.83-5.67 (m, 1H), 5.61- 5.58 (m, 1H), 4.25-4.22 (m, 1H), 4.14-4.09 (m,1H), 3.32-3.29 (m, 1H), 3.21 (s, 3H), 2.06-2.04 (m, 1H), 1.59- 1.55 (m,1H), 1.47-1.44 (m, 1H), 1.28-1.25 (m, 1H) 771

[M + NH₄]⁺ 418 772

[M + NH₄]⁺ 393 773

[M + NH₄]⁺ 340 774

[M + NH₄]⁺ 378 775

[M + NH₄]⁺ 366 776

[M + NH₄]⁺ 260 777

[M + NH₄]⁺ 296 778

[M + NH₄]⁺ 310 779

[M + NH₄]⁺ 336 780

[M + NH₄]⁺ 408 781

[M + NH₄]⁺ 372 782

(400 MHz, CDCl₃): δ 7.89 (d, 1H), 6.98 (d, 1H), 6.55 (t, J = 54 Hz, 1H),5.41 (d, 1H), 4.68 (m, 1H), 3.98-3.88 (m, 2H), 3.66-3.54 (m, 2H),3.48-3.26 (m, 2H), 1.87-1.74 (m, 2H), 1.62-1.51 (m, 2H) 783

[M + NH₄]⁺ 402 784

[M + NH₄]⁺ 392 785

[M + NH₄]⁺ 406 786

[M + NH₄]⁺ 415 787

[M + NH₄]⁺ 415 788

[M + NH₄]⁺ 429 789

[M + NH₄]⁺ 429 790

[M + NH₄]⁺ 429 791

[M + HCl − H]⁻ 508 792

[M + H]⁺ 374 793

[M + NH₄]⁺ 469 794

[M + NH₄]⁺ 411 795

[M + NH₄]⁺ 411 796

[M + NH₄]⁺ 384 797

402 (M + H) 798

[M + NH₄]⁺ 404 799

[M + NH₄]⁺ 354 800

[M + NH₄]⁺ 350 801

(300 MHz, CDCl₃): δ 8.25 (d, 1H), 7.27 (d, 1H), 4.18 (t, 2H), 3.50 (t,2H), 1.94 (m, 2H), 1.11 (t, 3H) 802

(300 MHz, CDCl₃): δ 8.23 (d, 1H), 7.25 (d, 1H), 4.83 (m, 1H), 3.47 (t,2H), 1.48 (d, 6H) 803

804

805

(400 MHz, CDCl₃): δ 8.21 (d, 1H), 7.40 (d, 1H), 4.88-4.83 (m, 1H), 3.67-3.39 (m, 7H), 1.43 (d, 3H) 806

(400 MHz, CDCl₃): δ 8.13 (d, 1H), 7.34 (d, 1H), 4.82-4.76 (m, 1H), 3.60-3.31 (m, 7H), 1.36 (d, 3H) 807

405/407 (M + H) (400 MHz, CDCl₃): δ 8.38 (d, 1H), 7.37- 7.36 (m, 1H),7.30 (ddd, 1H), 7.24 (dt, 1H), 6.09 (dddd, 1H), 6.00 (dddd, 1H) 808

403/405 (M + H) (400 MHz, CDCl₃): δ 8.36 (d, 1H), 7.37- 7.35 (m, 1H),7.29 (ddd, 1H), 7.25 (dt, 1H), 5.86 (dd, 1H), 5.11 (ddd, 1H), 2.73 (d,1H) 809

385 (M + H) (400 MHz, CDCl₃): δ 8.75 (d, 1H), 7.39- 7.37 (m, 1H), 7.34(ddd, 1H), 7.29-7.25 (m, 1H), 6.25 (ddd, 1H), 5.94-5.87 (m, 1H), 5.44(ddd, 1H), 3.66-3.58 (m, 1H), 3.29 (s, 3H) 810

367 (M + H) (400 MHz, CDCl₃): δ 8.60-8.59 (m, 1H), 7.35-7.33 (m, 1H),7.31 (ddd, 1H), 7.24 (dt, 1H), 5.63 (ddd, 1H), 5.51 (dtd, 1H), 3.76 (dd,1H), 3.47-3.55 (m, 1H), 3.31 (s, 3H), 3.29-3.15 (m, 1H) 811

403 (M + H) (400 MHz, CDCl₃): δ 8.80 (d, 1H), 7.40- 7.38 (m, 1H), 7.36(ddd, 1H), 7.28 (dt, 1H), 5.88 (dd, 1H), 5.66-5.60 (m, 1H), 3.52 (dd,1H), 3.28 (s, 3H) 812

403 (M + H) (400 MHz, CDCl₃): δ 8.77 (d, 1H), 7.39- 7.37 (m, 1H), 7.35(ddd, 1H), 7.27 (dt, 1H), 6.00 (dd, 1H), 5.83 (tdd, 1H), 3.97 (d, 1H),3.29 (s, 3H) 813

375 (M + H) (400 MHz, CDCl₃): δ 8.42 (s, 1H), 7.28 (ddd, 1H), 7.20-7.18(m, 1H), 7.09 (dt, 1H), 5.92 (dt, 1H), 5.53-5.46 (m, 1H), 5.19 (ddt,1H), 2.66 (ddd, 1H) 814

298 (M + H) 815

298 (M + H) 816

280 (M + H) 817

280 (M + H) 818

302 (M + H) 819

349 (M + H) (400 MHz, CDCl₃): δ 8.52-8.51 (m, 1H), 7.33 (ddd, 1H), 7.29(ddd, 1H), 7.23 (dt, 1H), 5.71-5.66 (m, 1H), 3.64 (d, 1H), 3.26-3.17 (m,1H), 3.22 (s, 3H), 2.99- 2.90 (ddd, 1H), 2.66-2.56 (m, 1H), 2.32- 2.22(m, 1H) 820

367 (M + H) (400 MHz, CDCl₃): δ 8.60-8.59 (m, 1H), 7.35-7.33 (m, 1H),7.31 (ddd, 1H), 7.24 (dt, 1H), 5.63 (ddd, 1H), 5.52 (dtd, 1H), 3.76 (dd,1H), 3.47-3.35 (m, 1H), 3.31 (s, 3H), 3.29-3.15 (m, 1H) 821

310 (M + H) 822

310 (M + H) 823

339 (M + H) (400 MHz, CDCl₃): δ 8.29-8.27 (m, 1H), 7.34-7.32 (m, 1H),7.26 (ddd, 1H), 7.22 (dt, 1H), 5.57-5.10 (m, 1H), 3.22 (dt, 1H), 2.96(ddd, 1H), 2.56-2.45 (m, 1H), 2.33-2.25 (m, 1H), 2.22-2.18 (m, 1H) 824

357 (M + H) (400 MHz, CDCl₃): δ 8.36-8.34 (m, 1H), 7.35-7.32 (m, 1H),7.28 (ddd, 1H), 7.23 (dt, 1H), 5.53-5.35 (m, 2H), 3.40 (ddd, 1H), 3.22(ddd, 1H), 2.73-2.68 (m, 1H) 825

358 (M + H) (400 MHz, CDCl₃): δ 8.79 (dd, 1H), 8.71 (d, 1H), 8.41 (s,1H), 7.67 (dd, 1H), 5.94 (dt, 1H), 5.53-5.46 (m, 1H), 5.21 (ddt, 1H),2.73 (ddd, 1H) 826

298 (M + H) (400 MHz, CDCl₃): δ 8.68 (s, 1H), 7.55- 7.45 (m, 3H),7.44-7.40 (m, 2H), 5.54- 5.46 (m, 1H), 5.31 (ddt, 1H), 3.46 (ddd, 1H),3.24 (ddd, 1H), 2.61 (dd, 1H) 827

334 (M + H) (400 MHz, CDCl₃): δ 8.65 (s, 1H), 6.99- 6.91 (m, 3H),5.54-5.47 (m, 1H), 5.33 (ddt, 1H), 3.44 (ddd, 1H), 3.23 (ddd, 1H), 2.63(dd, 1H) 828

346 (M + H) (400 MHz, CDCl₃): δ 8.24 (s, 1H), 5.95 (ddd, 1H), 5.47-5.41(m, 1H), 5.07 (ddt, 1H), 4.97-4.88 (m, 1H), 3.30-3.15 (m, 2H), 2.99-2.79(m, 2H), 2.54-2.49 (m, 1H) 829

315 (M + H) (400 MHz, DMSO-d₆): δ 8.30 (s, 1H), 6.26 (tt, 1H), 6.15(ddd, 1H), 4.55 (td, 2H), 3.46-3.32 (m, 1H), 3.13 (ddd, 1H) 830

324 (M + H) (400 MHz, CDCl₃): δ 8.14 (s, 1H), 6.10 (tt, 1H), 5.58-5.30(m, 1H), 4.39-4.24 (m, 2H), 3.10 (dt, 1H), 2.87 (ddd, 1H), 2.37-2.27 (m,1H), 2.20-2.12 (m, 1H), 1.90-1.86 (m, 1H), 1.47-1.39 (m, 2H), 1.27-1.21(m, 2H) 831

351 (M + H) 832

351 (M + H) (400 MHz, CDCl₃): δ 8.75 (d, 1H), 8.65- 8.62 (m, 1H), 6.17(tt, 1H), 5.83 (dd, 1H), 4.83-4.64 (m, 2H) 833

340 (M + H) (400 MHz, CDCl₃): δ 7.43-7.41 (m, 1H), 7.33 (dt, 1H),7.33-7.29 (m, 1H), 5.67- 5.62 (m, 1H), 3.36-3.26 (m, 1H), 3.05 (ddd,1H), 2.63-2.53 (m, 1H), 2.40-2.30 (m, 2H)

In some embodiments, the invention provides a combination treatmentcomprising a HIF-2α inhibitor, which can be a compound as providedherein, and an immunotherapeutic agent also as provided herein. In someembodiments, the immunotherapeutic agent is nivolumab (BMS936558),pembrolizumab (MK-3475), pidilizumab (CT-011), AMP-224, AMP-514,BMS-936559, RG7446 (MPDL3280A), MDX-1106 (Medarex Inc.), MSB0010718C,MEDI4736, tremelimumab or ipilimumab. In some embodiments, the HIF-2αinhibitor is a compound of Formula I, I-A, I-B, I-C, I-D, I-E, I-F, I-G,I-H, I-I, I-J, I-K, Formula II, II-A, II-B or a compound in Table 1, andthe immunotherapeutic agent is a PD-1 inhibitor or a CTLA-4 inhibitor.For example, the HIF-2α inhibitor is a compound of Formula I-C, wherein:X is CR⁵; Y is CR⁶; Z is —O—; R¹ is phenyl, optionally substituted withone or more substituents selected from the group consisting of fluoroand cyano; R⁴ is sulfonyl; R¹¹ is hydroxy; R¹² is hydrogen; R¹³ isfluoro; n=1 or 2; and R⁵ and R⁶ are hydrogen; and the immunotherapeuticagent is an agent that inhibits the PD-1 pathway and enhances an immuneresponse to cancerous cells in a patient. In a further embodiment, thePD-1 inhibitor is an antibody or a small molecule. In anotherembodiment, the immunotherapeutic agent is an agent that inhibits theCTLA-4 pathway and enhances an immune response to cancerous cells in apatient. In some embodiments, the CTLA-4 inhibitor is an antibody or asmall molecule. In another embodiment, the immunotherapeutic agent is anagent that inhibits a B7 protein and enhances an immune response tocancerous cells in patient.

In some other embodiments, the subject methods are useful for treating adisease condition associated with HIF-2, PD-1, CTLA-4 and/or a B7protein. Any disease condition characterized by an abnormal activity orexpression level of HIF-2α can be an intended disease condition. In someembodiments, the disease condition is a proliferative disorder, such asdescribed herein, including but not limited to cancer. A role of HIF-2αin tumorigenesis and tumor progression has been implicated in many humancancers. Constitutively active HIF-2α may be the result of defective VHLor a low concentration of oxygen in a cancer cell. Rapidly growingtumors are normally hypoxic due to poor vascularization, a conditionthat activates HIF-2α in support of tumor cell survival andproliferation. Constitutive activation of HIF-2α is emerging as a commontheme in diverse human cancers, consequently agents that target HIF-2αhave therapeutic value.

In still other embodiments, any disease condition in which cells of theafflicted subject express PD-L1 can be an intended disease condition.The subject methods are particularly suitable for treating a subjecthaving a disorder that can be treated by potentiating a T-cell mediatedimmune response. In some embodiments, the disorder is a proliferativedisorder, such as described herein, including but not limited to cancer.

Where desired, the subject to be treated is tested prior to treatmentusing a diagnostic assay to determine the sensitivity of tumor cells toa HIF-2α inhibitor or an immunotherapeutic agent. Any method known inthe art that can determine the sensitivity of the tumor cells of asubject to a HIF-2α inhibitor or an immunotherapeutic agent can beemployed. In one embodiment, when the subject is identified as one whosetumor cells are predicted to have high sensitivity to a HIF-2α inhibitoras a single agent, but may also display enhanced sensitivity in thepresence of a PD-1 inhibitor based on the results of a diagnostic assay,the subject is administered a therapeutically effective amount of acombination of a HIF-2α inhibitor and a PD-1 inhibitor. In anotherembodiment, when detection of PD-L1 expression in a tumor of the subjectpredicts for efficacy of a PD-1 inhibitor as a single agent, but mayalso display enhanced sensitivity in the presence of a HIF-2α inhibitorbased on the results of a diagnostic assay, the subject is administereda therapeutically effective amount of a combination of a HIF-2αinhibitor and a PD-1 inhibitor. In another embodiment, when detection ofCTLA-4 expression in a tumor of the subject predicts for efficacy of aCTLA-4 inhibitor as a single agent, but may also display enhancedsensitivity in the presence of a HIF-2α inhibitor based on the resultsof a diagnostic assay, the subject is administered a therapeuticallyeffective amount of a combination of a HIF-2α inhibitor and a CTLA-4inhibitor. In these methods, one or more additional anti-cancer agentsor treatments can be co-administered simultaneously or sequentially withthe HIF-2α inhibitor and the immunotherapeutic agent, as judged to beappropriate by the administering physician given the prediction of thelikely responsiveness of the subject to the combination of HIF-2αinhibitor and immunotherapeutic agent, in combination with anyadditional circumstances pertaining to the individual subject.

The data presented in the Examples herein demonstrate that theanti-tumor effects of a combination of a HIF-2α inhibitor and a PD-1inhibitor are superior to the anti-tumor effects of either inhibitor byitself. As such, the subject method is particularly useful for treatinga proliferative disorder, such as a neoplastic condition. Non-limitingexamples of such conditions include but are not limited to acanthoma,acinic cell carcinoma, acoustic neuroma, acral lentiginous melanoma,acrospiroma, acute eosinophilic leukemia, acute lymphoblastic leukemia,acute megakaryoblastic leukemia, acute monocytic leukemia, acutemyeloblastic leukemia with maturation, acute myeloid dendritic cellleukemia, acute myeloid leukemia, acute promyelocytic leukemia,adamantinoma, adenocarcinoma, adenoid cystic carcinoma, adenoma,adenomatoid odontogenic tumor, adrenocortical carcinoma, adult T-cellleukemia, aggressive NK-cell leukemia, AIDS-related cancers,AIDS-related lymphoma, alveolar soft part sarcoma, ameloblastic fibroma,anal cancer, anaplastic large cell lymphoma, anaplastic thyroid cancer,angioimmunoblastic T-cell lymphoma, angiomyolipoma, angiosarcoma,appendix cancer, astrocytoma, atypical teratoid rhabdoid tumor, basalcell carcinoma, basal-like carcinoma, B-cell leukemia, B-cell lymphoma,bellini duct carcinoma, biliary tract cancer, bladder cancer, blastoma,bone cancer, bone tumor, brain stem glioma, brain tumor, breast cancer,brenner tumor, bronchial tumor, bronchioloalveolar carcinoma, browntumor, Burkitt's lymphoma, carcinoid tumor, carcinoma, carcinosarcoma,Castleman's disease, central nervous system embryonal tumor, cerebellarastrocytoma, cerebral astrocytoma, cervical cancer, cholangiocarcinoma,chondroma, chondrosarcoma, chordoma, choriocarcinoma, choroid plexuspapilloma, chronic lymphocytic leukemia, chronic monocytic leukemia,chronic myelogenous leukemia, chronic myeloproliferative disorder,chronic neutrophilic leukemia, clear cell renal cell carcinoma,clear-cell tumor, colon cancer, colorectal cancer, craniopharyngioma,cutaneous T-cell lymphoma, dermatofibrosarcoma protuberans, dermoidcyst, desmoplastic small round cell tumor, diffuse large B celllymphoma, dysembryoplastic neuroepithelial tumor, embryonal carcinoma,endodermal sinus tumor, endometrial cancer, endometrial uterine cancer,endometrioid tumor, enteropathy-associated T-cell lymphoma,ependymoblastoma, ependymoma, epithelioid sarcoma, erythroleukemia,esophageal cancer, esthesioneuroblastoma, Ewing's sarcoma, extracranialgerm cell tumor, extragonadal germ cell tumor, extrahepatic bile ductcancer, extramammary Paget's disease, fallopian tube cancer, fibroma,fibrosarcoma, follicular lymphoma, follicular thyroid cancer,gallbladder cancer, ganglioglioma, ganglioneuroma, gastric cancer,gastric lymphoma, gastrointestinal cancer, gastrointestinal carcinoidtumor, gastrointestinal stromal tumor, germ cell tumor, germinoma,gestational choriocarcinoma, gestational trophoblastic tumor, giant celltumor of bone, glioblastoma multiforme, glioma, gliomatosis cerebri,glomus tumor, glucagonoma, gonadoblastoma, granulosa cell tumor, hairycell leukemia, head and neck cancer, heart cancer, hemangioblastoma,hemangiopericytoma, hemangiosarcoma, hematological malignancy,hepatocellular carcinoma, hepatosplenic T-cell lymphoma, Hodgkinlymphoma, hypopharyngeal cancer, hypothalamic glioma, inflammatorybreast cancer, intraocular melanoma, islet cell carcinoma, juvenilemyelomonocytic leukemia, Kaposi's sarcoma, kidney cancer, klatskintumor, krukenberg tumor, laryngeal cancer, lentigo maligna melanoma,leukemia, lip and oral cavity cancer, liposarcoma, lung cancer, luteoma,lymphangioma, lymphangiosarcoma, lymphoepithelioma, lymphoid leukemia,lymphoma, macroglobulinemia, malignant fibrous histiocytoma, malignantglioma, malignant mesothelioma, malignant peripheral nerve sheath tumor,malignant rhabdoid tumor, malignant triton tumor, malt lymphoma, mantlecell lymphoma, mast cell leukemia, mediastinal germ cell tumor,mediastinal tumor, medullary thyroid cancer, medulloblastoma,medulloepithelioma, melanoma, meningioma, merkel cell carcinoma,mesothelioma, metastatic squamous neck cancer with occult primary,metastatic urothelial carcinoma, mixed mullerian tumor, monocyticleukemia, mouth cancer, mucinous tumor, multiple endocrine neoplasiasyndrome, multiple myeloma, mycosis fungoides, myelodysplastic disease,myeloid leukemia, myeloid sarcoma, myeloproliferative disease, myxoma,nasal cavity cancer, nasopharyngeal cancer, neoplasm, neurinoma,neuroblastoma, neurofibroma, neuroma, nodular melanoma, non-Hodgkinlymphoma, nonmelanoma skin cancer, non-small cell lung cancer, ocularoncology, oligoastrocytoma, oligodendroglioma, oncocytoma, optic nervesheath meningioma, oral cancer, oropharyngeal cancer, osteosarcoma,ovarian cancer, ovarian epithelial cancer, ovarian germ cell tumor,ovarian low malignant potential tumor, pancoast tumor, pancreaticcancer, papillary thyroid cancer, papillomatosis, paraganglioma,paranasal sinus cancer, parathyroid cancer, penile cancer, perivascularepithelioid cell tumor, pharyngeal cancer, pheochromocytoma, pinealparenchymal tumor of intermediate differentiation, pineoblastoma,pituicytoma, pituitary adenoma, pituitary tumor, plasma cell neoplasm,pleuropulmonary blastoma, polyembryoma, precursor T-lymphoblasticlymphoma, primitive neuroectodermal tumor, prostate cancer, pseudomyxomaperitonei, rectal cancer, renal cell carcinoma, retinoblastoma,rhabdomyoma, rhabdomyosarcoma, Richter's transformation, sacrococcygealteratoma, salivary gland cancer, sarcoma, schwannomatosis, sebaceousgland carcinoma, secondary neoplasm, seminoma, serous tumor,Sertoli-Leydig cell tumor, sex cord-stromal tumor, sezary syndrome,signet ring cell carcinoma, skin cancer, small blue round cell tumor,small cell carcinoma, small cell lung cancer, small cell lymphoma, smallintestine cancer, soft tissue sarcoma, somatostatinoma, soot wart,spinal tumor, splenic marginal zone lymphoma, squamous cell carcinoma,stomach cancer, superficial spreading melanoma, supratentorial primitiveneuroectodermal tumor, surface epithelial-stromal tumor, synovialsarcoma, T-cell acute lymphoblastic leukemia, T-cell large granularlymphocyte leukemia, T-cell leukemia, T-cell lymphoma, T-cellprolymphocytic leukemia, teratoma, terminal lymphatic cancer, testicularcancer, thecoma, throat cancer, thymic carcinoma, thymoma, thyroidcancer, transitional cell cancer of renal pelvis and ureter,transitional cell carcinoma, urachal cancer, urethral cancer, urogenitalneoplasm, uterine sarcoma, uveal melanoma, vaginal cancer, vernermorrison syndrome, verrucous carcinoma, visual pathway glioma, vulvarcancer, Waldenstrom's macroglobulinemia, Warthin's tumor, Wilms' tumoror any combination thereof.

In some embodiments, the methods comprising administering a HIF-2αinhibitor and an immunotherapeutic agent described herein are applied tothe treatment of cancers of the adrenal glands, blood, bone marrow,brain, breast, cervix, colon, head and neck, kidney, liver, lung, ovary,pancreas, plasma cells, rectum, retina, skin, spine, throat, or anycombination thereof.

Certain embodiments contemplate a human subject such as a subject thathas been diagnosed as having or being at risk for developing oracquiring a proliferative disorder condition. Certain other embodimentscontemplate a non-human subject, for example a non-human primate such asa macaque, chimpanzee, gorilla, vervet, orangutan, baboon or othernon-human primate, including such non-human subjects that can be knownto the art as preclinical models. Certain other embodiments contemplatea non-human subject that is a mammal, for example, a mouse, rat, rabbit,pig, sheep, horse, bovine, goat, gerbil, hamster, guinea pig or othermammal. There are also contemplated other embodiments in which thesubject or biological source can be a non-mammalian vertebrate, forexample, another higher vertebrate, or an avian, amphibian or reptilianspecies, or another subject or biological source. In certain embodimentsof the present invention, a transgenic animal is utilized. A transgenicanimal is a non-human animal in which one or more of the cells of theanimal includes a nucleic acid that is non-endogenous (i.e.,heterologous) and is present as an extrachromosomal element in a portionof its cell or stably integrated into its germ line DNA (i.e., in thegenomic sequence of most or all of its cells).

Therapeutic Efficacy:

In some embodiments, therapeutic efficacy is measured based on an effectof treating a proliferative disorder, such as cancer. In general,therapeutic efficacy of the methods and compositions of the invention,with regard to the treatment of a proliferative disorder (e.g. cancer,whether benign or malignant), may be measured by the degree to which themethods and compositions promote inhibition of tumor cell proliferation,the inhibition of tumor vascularization, the eradication of tumor cells,the reduction in the rate of growth of a tumor, and/or a reduction inthe size of at least one tumor. Several parameters to be considered inthe determination of therapeutic efficacy are discussed herein. Theproper combination of parameters for a particular situation can beestablished by the clinician. The progress of the inventive method intreating cancer (e.g., reducing tumor size or eradicating cancerouscells) can be ascertained using any suitable method, such as thosemethods currently used in the clinic to track tumor size and cancerprogress. The primary efficacy parameter used to evaluate the treatmentof cancer by the inventive method and compositions preferably is areduction in the size of a tumor. Tumor size can be figured using anysuitable technique, such as measurement of dimensions, or estimation oftumor volume using available computer software, such as FreeFlightsoftware developed at Wake Forest University that enables accurateestimation of tumor volume. Tumor size can be determined by tumorvisualization using, for example, CT, ultrasound, SPECT, spiral CT, MRI,photographs, and the like. In embodiments where a tumor is surgicallyresected after completion of the therapeutic period, the presence oftumor tissue and tumor size can be determined by gross analysis of thetissue to be resected, and/or by pathological analysis of the resectedtissue.

In some desirable embodiments, the growth of a tumor is stabilized(i.e., one or more tumors do not increase more than 1%, 5%, 10%, 15%, or20% in size, and/or do not metastasize) as a result of the inventivemethod and compositions. In some embodiments, a tumor is stabilized forat least about 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, or more weeks. Insome embodiments, a tumor is stabilized for at least about 1, 2, 3, 4,5, 6, 7, 8, 9, 10, 11, 12, or more months. In some embodiments, a tumoris stabilized for at least about 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, or moreyears. Preferably, the inventive method reduces the size of a tumor atleast about 5% (e.g., at least about 10%, 15%, 20%, or 25%). Morepreferably, tumor size is reduced at least about 30% (e.g., at leastabout 35%, 40%, 45%, 50%, 55%, 60%, or 65%). Even more preferably, tumorsize is reduced at least about 70% (e.g., at least about 75%, 80%, 85%,90%, or 95%). Most preferably, the tumor is completely eliminated, orreduced below a level of detection. In some embodiments, a subjectremains tumor free (e.g. in remission) for at least about 1, 2, 3, 4, 5,6, 7, 8, 9, 10, 11, 12, or more weeks following treatment. In someembodiments, a subject remains tumor free for at least about 1, 2, 3, 4,5, 6, 7, 8, 9, 10, 11, 12, or more months following treatment. In someembodiments, a subject remains tumor free for at least about 1, 2, 3, 4,5, 6, 7, 8, 9, 10, or more years after treatment.

In some embodiments, the efficacy of the inventive method in reducingtumor size can be determined by measuring the percentage of necrotic(i.e., dead) tissue of a surgically resected tumor following completionof the therapeutic period. In some further embodiments, a treatment istherapeutically effective if the necrosis percentage of the resectedtissue is greater than about 20% (e.g., at least about 30%, 40%, 50%,60%, 70%, 80%, 90%, or 100%), more preferably about 90% or greater(e.g., about 90%, 95%, or 100%). Most preferably, the necrosispercentage of the resected tissue is 100%, that is, no tumor tissue ispresent or detectable.

The efficacy of the inventive method can be determined by a number ofsecondary parameters. Examples of secondary parameters include, but arenot limited to, detection of new tumors, detection of tumor antigens ormarkers (e.g., CEA, PSA, or CA-125), biopsy, surgical downstaging (i.e.,conversion of the surgical stage of a tumor from unresectable toresectable), PET scans, survival, disease progression-free survival,time to disease progression, quality of life assessments such as theClinical Benefit Response Assessment, and the like, all of which canpoint to the overall progression (or regression) of cancer in a human.Biopsy is particularly useful in detecting the eradication of cancerouscells within a tissue. Radioimmunodetection (RAID) is used to locate andstage tumors using serum levels of markers (antigens) produced by and/orassociated with tumors (“tumor markers” or “tumor-associated antigens”),and can be useful as a pre-treatment diagnostic predicate, apost-treatment diagnostic indicator of recurrence, and a post-treatmentindicator of therapeutic efficacy. Examples of tumor markers ortumor-associated antigens that can be evaluated as indicators oftherapeutic efficacy include, but are not limited to, carcinoembryonicantigen (CEA), prostate-specific antigen (PSA), CA-125, CA19-9,ganglioside molecules (e.g., GM2, GD2, and GD3), MART-1, heat shockproteins (e.g., gp96), sialyl Tn (STn), tyrosinase, MUC-1, HER-2/neu,c-erb-B2, KSA, PSMA, p53, RAS, EGF-R, VEGF, MAGE, and gp100. Othertumor-associated antigens are known in the art. RAID technology incombination with endoscopic detection systems also can efficientlydistinguish small tumors from surrounding tissue (see, for example, U.S.Pat. No. 4,932,412).

In additional desirable embodiments, the treatment of cancer in a humanpatient in accordance with the inventive method is evidenced by one ormore of the following results: (a) the complete disappearance of a tumor(i.e., a complete response), (b) about a 25% to about a 50% reduction inthe size of a tumor for at least four weeks after completion of thetherapeutic period as compared to the size of the tumor beforetreatment, (c) at least about a 50% reduction in the size of a tumor forat least four weeks after completion of the therapeutic period ascompared to the size of the tumor before the therapeutic period, and (d)at least a 2% decrease (e.g., about a 5%, 10%, 20%, 30%, 40%, 50%, 60%,70%, 80% or 90% decrease) in a specific tumor-associated antigen levelat about 4-12 weeks after completion of the therapeutic period ascompared to the tumor-associated antigen level before the therapeuticperiod. While at least a 2% decrease in a tumor-associated antigen levelis preferred, any decrease in the tumor-associated antigen level isevidence of treatment of a cancer in a patient by the inventive method.For example, with respect to unresectable, locally advanced pancreaticcancer, treatment can be evidenced by at least a 10% decrease in theCA19-9 tumor-associated antigen level at 4-12 weeks after completion ofthe therapeutic period as compared to the CA19-9 level before thetherapeutic period. Similarly, with respect to locally advanced rectalcancer, treatment can be evidenced by at least a 10% decrease in the CEAtumor-associated antigen level at 4-12 weeks after completion of thetherapeutic period as compared to the CEA level before the therapeuticperiod.

With respect to quality of life assessments, such as the ClinicalBenefit Response Criteria, the therapeutic benefit of the treatment inaccordance with the invention can be evidenced in terms of painintensity, analgesic consumption, and/or the Karnofsky Performance Scalescore. The treatment of cancer in a human patient alternatively, or inaddition, is evidenced by (a) at least a 50% decrease (e.g., at least a60%, 70%, 80%, 90%, or 100% decrease) in pain intensity reported by apatient, such as for any consecutive four week period in the 12 weeksafter completion of treatment, as compared to the pain intensityreported by the patient before treatment, (b) at least a 50% decrease(e.g., at least a 60%, 70%, 80%, 90%, or 100% decrease) in analgesicconsumption reported by a patient, such as for any consecutive four weekperiod in the 12 weeks after completion of treatment as compared to theanalgesic consumption reported by the patient before treatment, and/or(c) at least a 20 point increase (e.g., at least a 30 point, 50 point,70 point, or 90 point increase) in the Karnofsky Performance Scale scorereported by a patient, such as for any consecutive four week period inthe 12 weeks after completion of the therapeutic period as compared tothe Karnofsky Performance Scale score reported by the patient before thetherapeutic period.

In some cases, conventional response criteria may not adequately assessthe activity of immunotherapeutic agents (i.e., a PD-1 inhibitor)because progressive disease (by initial radiographic evaluation) doesnot necessarily reflect therapeutic failure. Accordingly, to properlyevaluate immunotherapeutic agents, long-term effects on the targetdisease may also be captured. In this regard, systemic immune-relatedresponse criteria (irRC) that make allowances for an early increase intumor burden and/or the appearance of new lesions, and which seek toenhance the characterization of immune-related response patterns, havebeen proposed (see e.g. Wolchok et al., 2009).

The treatment of a proliferative disorder (e.g. cancer, whether benignor malignant) in a human patient desirably is evidenced by one or more(in any combination) of the foregoing results, although alternative oradditional results of the referenced tests and/or other tests canevidence treatment efficacy.

In some embodiments, tumor size is reduced as a result of the inventivemethod preferably without significant adverse events in the subject.Adverse events are categorized or “graded” by the Cancer TherapyEvaluation Program (CTEP) of the National Cancer Institute (NCI), withGrade 0 representing minimal adverse side effects and Grade 4representing the most severe adverse events. Desirably, the inventivemethod is associated with minimal adverse events, e.g. Grade 0, Grade 1,or Grade 2 adverse events, as graded by the CTEP/NCI. However, asdiscussed herein, reduction of tumor size, although preferred, is notrequired in that the actual size of tumor may not shrink despite theeradication of tumor cells. Eradication of cancerous cells is sufficientto realize a therapeutic effect. Likewise, any reduction in tumor sizeis sufficient to realize a therapeutic effect.

Detection, monitoring and rating of various cancers in a human arefurther described in Cancer Facts and Figures 2001, American CancerSociety, New York, N.Y., and International Patent Application WO01/24684. Accordingly, a clinician can use standard tests to determinethe efficacy of the various embodiments of the inventive method intreating cancer. However, in addition to tumor size and spread, theclinician also may consider quality of life and survival of the patientin evaluating efficacy of treatment.

In some embodiments, administration of a HIF-2α inhibitor and animmunotherapeutic agent provides improved therapeutic efficacy overtreatment with either agent alone. Improved efficacy may be measuredusing any method known in the art, including but not limited to thosedescribed herein. In some embodiments, the improved therapeutic efficacyis an improvement of at least about 10%, 20%, 30%, 40%, 50%, 60%, 70%,75%, 80%, 90%, 95%, 100%, 110%, 120%, 150%, 200%, 300%, 400%, 500%,600%, 700%, 1000% or more, using an appropriate measure (e.g. tumor sizereduction, duration of tumor size stability, duration of time free frommetastatic events, duration of disease-free survival). Improved efficacymay also be expressed as fold improvement, such as at least about2-fold, 3-fold, 4-fold, 5-fold, 6-fold, 7-fold, 8-fold, 9-fold, 10-fold,20-fold, 30-fold, 40-fold, 50-fold, 60-fold, 70-fold, 80-fold, 90-fold,100-fold, 1000-fold, 10000-fold or more, using an appropriate measure(e.g. tumor size reduction, duration of tumor size stability, durationof time free from metastatic events, duration of disease-free survival).

In some other embodiments, the subject method further comprisesmeasuring a tumor size or an amount of one or more markers for thepresence of the proliferative disorder before and after administeringthe HIF-2α inhibitor and the immunotherapeutic agent, and adjustingdosage or discontinuing use of the HIF-2α inhibitor and/or theimmunotherapeutic agent based on results of said measuring.

Pharmaceutical Compositions:

The invention provides, in one aspect, a combination treatmentcomprising administering to a subject a HIF-2α inhibitor and animmunotherapeutic agent. The HIF-2α inhibitor can be any HIF-2αinhibitor described herein, either alone or in combination with one ormore other such HIF-2α inhibitors. The immunotherapeutic agent can beany immunotherapeutic agent described herein, either alone or incombination with one or more other immunotherapeutic agents. In someembodiments, the HIF-2α inhibitor and the immunotherapeutic agent areadministered sequentially. In some further embodiments, theimmunotherapeutic agent is administered after administration of theHIF-2α inhibitor. In still further embodiments, the HIF-2α inhibitor isadministered after administration of the immunotherapeutic agent. In thesequential administration protocol, a HIF-2α inhibitor and animmunotherapeutic agent may be administered a few hours apart, a fewdays apart, a few weeks apart, or a few months apart. In some otherembodiments, the HIF-2α inhibitor and the immunotherapeutic agent areadministered simultaneously. In a further embodiment, the HIF-2αinhibitor and the immunotherapeutic agent form part of the samecomposition. In still a further embodiment, the HIF-2α inhibitor and theimmunotherapeutic agent are provided in one or more unit doses. In someembodiments, a composition comprising both a HIF-2α inhibitor and animmunotherapeutic agent releases the majority of the immunotherapeuticagent (e.g. at least 60%, 70%, 80%, 85%, 90%, 95%, 99%, or more) as anactive compound after the release of the majority of the HIF-2αinhibitor (e.g. at least 60%, 70%, 80%, 85%, 90%, 95%, 99%, or more) asan active compound. In some embodiments, a composition comprising both aHIF-2α inhibitor and an immunotherapeutic agent releases the majority ofthe HIF-2α inhibitor (e.g. at least 60%, 70%, 80%, 85%, 90%, 95%, 99%,or more) as an active compound after the release of the majority of theimmunotherapeutic agent (e.g. at least 60%, 70%, 80%, 85%, 90%, 95%,99%, or more) as an active compound.

In some embodiments, a HIF-2α inhibitor and/or an immunotherapeuticagent is administered to a subject more than once. In some embodiments,a HIF-2α inhibitor is administered one or more times (e.g. 1, 2, 3, 4,5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 20, 25, or more) every 1, 2, 3,4, 5, 6, 7, or more days (e.g. daily, every other day, every 7 days),where one or more of the administrations of the HIF-2α inhibitor isaccompanied by or followed by one or more administrations (e.g. 1, 2, 3,4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 20, 25, or more) of animmunotherapeutic agent with any desired temporal spacing. In someembodiments, a HIF-2α inhibitor is administered one or more times (e.g.1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 20, 25, or more)every 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, or more weeks (e.g.administration on 1, 2, 3, 4, 5, 6, and/or 7 days of a week, which mayor may not be consecutive days), where one or more of theadministrations of the HIF-2α inhibitor is accompanied by or followed byone or more administrations (e.g. 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12,13, 14, 15, 20, 25, or more) of an immunotherapeutic agent with anydesired temporal spacing. In some embodiments, a given dosing schedulecomprising one or more administrations of a HIF-2α inhibitor and one ormore administrations of an immunotherapeutic agent may be repeated on adaily, weekly, biweekly, monthly, bimonthly, annually, semi-annually, orany other period as may be determined by a medical professional. Arepeated dosing schedule may be repeated for a fixed period of timedetermined at the start of the schedule; may be terminated, extended, orotherwise adjusted based on a measure of therapeutic effect, such as alevel of reduction in the presence of detectable disease tissue (e.g. areduction of at least 50%, 60%, 70%, 80%, 90%, 95%, 99%, or 100%); ormay be terminated, extended, or otherwise adjusted for any other reasonas determined by a medical professional.

A combination treatment may further comprise the administration of oneor more additional therapeutic agents, including one or more additionalagents described herein as candidate HIF-2α inhibitors, and one or moreadditional agents described herein as candidate immunotherapeuticagents. Such one or more additional agents can be administeredsimultaneously or separately with respect to the HIF-2α inhibitor, theimmunotherapeutic agent, or both. Administration in combinationutilizing one or more additional agents includes, for example,simultaneous administration of two agents in the same dosage form,simultaneous administration in separate dosage forms, and separateadministration. For example, multiple therapeutic agents can beformulated together in the same dosage form and administeredsimultaneously. Alternatively, multiple therapeutic agents can besimultaneously administered, wherein both the agents are present inseparate formulations. In another alternative, an inhibitor of thepresent invention can be administered after any of the agents describedabove, or vice versa. In the separate administration protocol, aninhibitor of the present invention and any of the agents described abovemay be administered a few minutes apart, or a few hours apart, or a fewdays apart, or a few weeks apart. The term “combination treatments” alsoembraces the administration of the therapeutic agents as describedherein in further combination with other biologically active compoundsor ingredients and non-drug therapies (e.g., surgery or radiationtreatment).

Administration of the compounds of the present invention can proceed byany method that enables delivery of the compounds to the site of action.These methods include oral routes, intraduodenal routes, parenteralinjection (including intravenous, intraarterial, subcutaneous,intramuscular, intravascular, intraperitoneal or infusion), topical(e.g., transdermal application), rectal administration, via localdelivery by catheter or stent or through inhalation. Compounds can alsobe administered intraadiposally or intrathecally. An effective amount ofan inhibitor of the invention may be administered in either single ormultiple doses by any of the accepted modes of administration of agentshaving similar utilities, including rectal, buccal, intranasal andtransdermal routes, by intra-arterial injection, intravenously,intraperitoneally, parenterally, intramuscularly, subcutaneously,orally, topically, as an inhalant, or via an impregnated or coateddevice such as a stent, for example, or an artery-inserted cylindricalpolymer. Sequential or simultaneous administration of a HIF-2αinhibitor, an immunotherapeutic agent, and/or any additional therapeuticagent can be effected by any appropriate route as noted above andincluding, but not limited to, oral routes, intravenous routes,intramuscular routes, and direct absorption through mucous membranetissues. The therapeutic agents can be administered by the same route orby different routes. For example, a first therapeutic agent of thecombination selected may be administered by intravenous injection whilethe other therapeutic agents of the combination may be administeredorally. Alternatively, for example, all therapeutic agents may beadministered orally or all therapeutic agents may be administered byintravenous injection.

Methods of determining the most effective means and dosage ofadministration are well known to those of skill in the art and will varywith the composition used for therapy, the purpose of the therapy, thetarget cell or tissue being treated, and the subject being treated.Single or multiple administrations (e.g. about or more than about 1, 2,3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 25, 30,or more doses) can be carried out with the dose level and pattern beingselected by the treating physician.

A composition of the present disclosure may be formulated in anysuitable pharmaceutical formulation. A pharmaceutical composition of thepresent disclosure typically contains an active ingredient (e.g., acompound of the present disclosure or a pharmaceutically acceptable saltand/or coordination complex thereof), and one or more pharmaceuticallyacceptable excipients, carriers, including but not limited to, inertsolid diluents and fillers, diluents, sterile aqueous solution andvarious organic solvents, permeation enhancers, solubilizers andadjuvants. A composition of the present disclosure may be formulated inany suitable pharmaceutical formulation. In some embodiments, thepharmaceutical acceptable carriers, excipients are selected from thegroup consisting of water, alcohol, glycerol, chitosan, alginate,chondroitin, Vitamin E, mineral oil, and dimethyl sulfoxide (DMSO).

Pharmaceutical formulations may be provided in any suitable form, whichmay depend on the route of administration. In some embodiments, thepharmaceutical composition disclosed herein can be formulated in dosageform for administration to a subject. In some embodiments, thepharmaceutical composition is formulated for oral, intravenous,intraarterial, aerosol, parenteral, buccal, topical, transdermal,rectal, intramuscular, subcutaneous, intraosseous, intranasal,intrapulmonary, transmucosal, inhalation, and/or intraperitonealadministration. In some embodiments, the dosage form is formulated fororal intervention administration. For example, the pharmaceuticalcomposition can be formulated in the form of a pill, a tablet, acapsule, an inhaler, a liquid suspension, a liquid emulsion, a gel, or apowder. In some embodiments, the pharmaceutical composition can beformulated as a unit dosage in liquid, gel, semi-liquid, semi-solid, orsolid form.

The amount of each compound administered will be dependent on the mammalbeing treated, the severity of the disorder or condition, the rate ofadministration, the disposition of the compound and the discretion ofthe prescribing physician. However, an effective dosage may be in therange of about 0.001 to about 100 mg per kg body weight per day, insingle or divided doses. In some instances, dosage levels below thelower limit of the aforesaid range may be more than adequate, while inother cases still larger doses may be employed without causing anyharmful side effect, e.g., by dividing such larger doses into severalsmall doses for administration throughout the day.

In some embodiments, the disclosure provides a pharmaceuticalcomposition comprising an amount of a HIF-2α inhibitor formulated foradministration to a subject in need thereof. In some embodiments, thepharmaceutical composition comprises between about 0.0001-500 g,0.001-250 g, 0.1-50 g, or 1-10 g of a HIF-2α inhibitor. In someembodiments, the pharmaceutical composition comprises about or more thanabout 0.0001 g, 0.001 g, 0.01 g, 0.1, 0.5 g, 1 g, 2 g, 3 g, 4 g, 5 g, 6g, 7 g, 8 g, 9 g, 10 g, 15 g, 20 g, 25 g, 50 g, 2.0 g, 100 g, 200 g, 250g, 300 g, 350 g, 400 g, 450 g, 500 g or more of a HIF-2α inhibitor. Insome embodiments, the pharmaceutical composition comprises between0.001-2 g of a HIF-2α inhibitor in a single dose. In some embodiments,the pharmaceutical composition comprises an amount between about 50-150g of a HIF-2α inhibitor.

In some embodiments, a therapeutically effective amount of HIF-2αinhibitor, typically a daily amount administered over the course of aperiod of treatment, can sufficiently provide any one or more of thetherapeutic effects described herein. As an example, the therapeuticeffect amount can be found in the range of about 0.001-1000 mg/kg bodyweight, 0.001-50 mg/kg body weight, 0.01-100 mg/kg body weight, 0.01-30mg/kg body weight, 0.1-200 mg/kg body weight, 3-200 mg/kg body weight,5-500 mg/kg body weight, 10-100 mg/kg body weight, 10-1000 mg/kg bodyweight, 50-200 mg/kg body weight, 100-1000 mg/kg body weight, 200-500mg/kg body weight, 250-350 mg/kg body weight, or 300-600 mg/kg bodyweight of a HIF-2α inhibitor. In some embodiments, the therapeuticamount can be about or more than about 0.001 mg/kg body weight, 0.01mg/kg body weight, 0.1 mg/kg body weight, 0.5 mg/kg body weight, 1 mg/kgbody weight, 2 mg/kg body weight, 3 mg/kg body weight, 4 mg/kg bodyweight, 5 mg/kg body weight, 6 mg/kg body weight, 7 mg/kg body weight, 8mg/kg body weight, 9 mg/kg body weight, 10 mg/kg body weight, 15 mg/kgbody weight, 20 mg/kg body weight, 25 mg/kg body weight, 50 mg/kg bodyweight, 100 mg/kg body weight, 200 mg/kg body weight, 250 mg/kg bodyweight, 300 mg/kg body weight, 350 mg/kg body weight, 400 mg/kg bodyweight, 450 mg/kg body weight, 500 mg/kg body weight, 600 mg/kg bodyweight, 800 mg/kg body weight, 1000 mg/kg body weight, or more of aHIF-2α inhibitor. In some embodiments, the effective amount is at leastabout 0.01 mg/kg body weight of a HIF-2α inhibitor. In some embodiments,the effective amount is an amount between about 0.01-30 mg/kg bodyweight of a HIF-2α inhibitor. In some embodiments, the therapeuticamount can be an amount between about 50-150 mg/kg body weight of aHIF-2α inhibitor.

In some embodiments, dosage regimens are adjusted to provide the optimumdesired response. For administration of an immunotherapeutic agent, thedosage ranges from about 0.0001 to about 100 mg/kg, usually from about0.001 to about 20 mg/kg, and more usually from about 0.01 to about 10mg/kg, of the subject's body weight. Preferably, the dosage ofimmunotherapeutic agent is within the range of 0.1-10 mg/kg body weight.For example, dosages can be about 0.1, 0.3, 1, 3, 5 or 10 mg/kg bodyweight, and more preferably about 0.3, 1, 3 or 10 mg/kg body weight.

In some embodiments, a HIF-2α inhibitor, an immunotherapeutic agentand/or any additional therapeutic compound of the invention is providedin one or more unit doses. For example, the composition can beadministered in 1, 2, 3, 4, 5, 6, 7, 14, 30, 60, or more doses. Suchamount can be administered each day, for example in individual dosesadministered once, twice, or three or more times a day. However, dosagesstated herein on a per day basis should not be construed to requireadministration of the daily dose each and every day. For example, if oneof the agents is provided in a suitably slow-release form, two or moredaily dosage amounts can be administered at a lower frequency, e.g., asa depot every second day to once a month or even longer. Most typicallyand conveniently for the subject, a HIF-2α inhibitor, animmunotherapeutic agent and/or any additional therapeutic compound canbe administered once a day, for example in the morning, in the eveningor during the day.

The unit doses can be administered simultaneously or sequentially. Thecomposition can be administered for an extended treatment period.Illustratively, the treatment period can be at least about one month,for example at least about 3 months, at least about 6 months or at leastabout 1 year. In some cases, administration can continue forsubstantially the remainder of the life of the subject.

When a combination treatment of the invention is administered as acomposition that comprises one or more compounds, and one compound has ashorter half-life than another compound, the unit dose forms may beadjusted accordingly.

In some embodiments, combination treatments of the invention are testedto estimate pharmacokinetic properties and expected side effect profile.Various assays are known in the art for this purpose. For example, oralavailability can be estimated during early stages of drug development byperforming a Caco-2 permeability assay. Further, oral pharmacokineticsin humans can be approximated by extrapolating from the results ofassays in mice, rats or monkey. In some embodiments, compounds of theinvention show good oral availability across multiple species oforganisms.

In some embodiments, a HIF-2α inhibitor and an immunotherapeutic agentcan be administered as part of a therapeutic regimen that comprisesadministering one or more other agents (e.g. 1, 2, 3, 4, 5, or moreother agents), either simultaneously or sequentially with the HIF-2αinhibitor and the immunotherapeutic agent. When administeredsequentially, the HIF-2α inhibitor and/or the immunotherapeutic agentmay be administered before or after the one or more other agents. Whenadministered simultaneously, the HIF-2α inhibitor, the immunotherapeuticagent and the one or more other agents may be administered by the sameroute (e.g. injections to the same location; tablets taken orally at thesame time), by a different route (e.g. a tablet taken orally whilereceiving an intravenous infusion), or as part of the same combination(e.g. a solution comprising a HIF-2α inhibitor, an immunotherapeuticagent and one or more other agents).

In some embodiments, the concentration of one or more of the compoundsprovided in the pharmaceutical compositions of the present invention isless than 100%, 90%, 80%, 70%, 60%, 50%, 40%, 30%, 20%, 19%, 18%, 17%,16%, 15%, 14%, 13%, 12%, 11%, 10%, 9%, 8%, 7%, 6%, 5%, 4%, 3%, 2%, 1%,0.5%, 0.4%, 0.3%, 0.2%, 0.1%, 0.09%, 0.08%, 0.07%, 0.06%, 0.05%, 0.04%,0.03%, 0.02%, 0.01%, 0.009%, 0.008%, 0.007%, 0.006%, 0.005%, 0.004%,0.003%, 0.002%, 0.001%, 0.0009%, 0.0008%, 0.0007%, 0.0006%, 0.0005%,0.0004%, 0.0003%, 0.0002%, or 0.0001% w/w, w/v or v/v.

In some embodiments, the concentration of one or more of the compoundsof the present invention is greater than 90%, 80%, 70%, 60%, 50%, 40%,30%, 20%, 19.75%, 19.50%, 19.25% 19%, 18.75%, 18.50%, 18.25% 18%,17.75%, 17.50%, 17.25% 17%, 16.75%, 16.50%, 16.25% 16%, 15.75%, 15.50%,15.25% 15%, 14.75%, 14.50%, 14.25% 14%, 13.75%, 13.50%, 13.25% 13%,12.75%, 12.50%, 12.25% 12%, 11.75%, 11.50%, 11.25% 11%, 10.75%, 10.50%,10.25% 10%, 9.75%, 9.50%, 9.25% 9%, 8.75%, 8.50%, 8.25% 8%, 7.75%,7.50%, 7.25% 7%, 6.75%, 6.50%, 6.25% 6%, 5.75%, 5.50%, 5.25% 5%, 4.75%,4.50%, 4.25%, 4%, 3.75%, 3.50%, 3.25%, 3%, 2.75%, 2.50%, 2.25%, 2%,1.75%, 1.50%, 125%, 1%, 0.5%, 0.4%, 0.3%, 0.2%, 0.1%, 0.09%, 0.08%,0.07%, 0.06%, 0.05%, 0.04%, 0.03%, 0.02%, 0.01%, 0.009%, 0.008%, 0.007%,0.006%, 0.005%, 0.004%, 0.003%, 0.002%, 0.001%, 0.0009%, 0.0008%,0.0007%, 0.0006%, 0.0005%, 0.0004%, 0.0003%, 0.0002%, or 0.0001% w/w,w/v, or v/v.

In some embodiments, the concentration of one or more of the compoundsof the present invention is in the range from approximately 0.0001% toapproximately 50%, approximately 0.001% to approximately 40%,approximately 0.01% to approximately 30%, approximately 0.02% toapproximately 29%, approximately 0.03% to approximately 28%,approximately 0.04% to approximately 27%, approximately 0.05% toapproximately 26%, approximately 0.06% to approximately 25%,approximately 0.07% to approximately 24%, approximately 0.08% toapproximately 23%, approximately 0.09% to approximately 22%,approximately 0.1% to approximately 21%, approximately 0.2% toapproximately 20%, approximately 0.3% to approximately 19%,approximately 0.4% to approximately 18%, approximately 0.5% toapproximately 17%, approximately 0.6% to approximately 16%,approximately 0.7% to approximately 15%, approximately 0.8% toapproximately 14%, approximately 0.9% to approximately 12%, orapproximately 1% to approximately 10% w/w, w/v or v/v.

In some embodiments, the concentration of one or more of the compoundsof the present invention is in the range from approximately 0.001% toapproximately 10%, approximately 0.01% to approximately 5%,approximately 0.02% to approximately 4.5%, approximately 0.03% toapproximately 4%, approximately 0.04% to approximately 3.5%,approximately 0.05% to approximately 3%, approximately 0.06% toapproximately 2.5%, approximately 0.07% to approximately 2%,approximately 0.08% to approximately 1.5%, approximately 0.09% toapproximately 1%, or approximately 0.1% to approximately 0.9% w/w, w/vor v/v.

In some embodiments, the amount of one or more of the compounds of thepresent invention is equal to or less than 10 g, 9.5 g, 9.0 g, 8.5 g,8.0 g, 7.5 g, 7.0 g, 6.5 g, 6.0 g, 5.5 g, 5.0 g, 4.5 g, 4.0 g, 3.5 g,3.0 g, 2.5 g, 2.0 g, 1.5 g, 1.0 g, 0.95 g, 0.9 g, 0.85 g, 0.8 g, 0.75 g,0.7 g, 0.65 g, 0.6 g, 0.55 g, 0.5 g, 0.45 g, 0.4 g, 0.35 g, 0.3 g, 0.25g, 0.2 g, 0.15 g, 0.1 g, 0.09 g, 0.08 g, 0.07 g, 0.06 g, 0.05 g, 0.04 g,0.03 g, 0.02 g, 0.01 g, 0.009 g, 0.008 g, 0.007 g, 0.006 g, 0.005 g,0.004 g, 0.003 g, 0.002 g, 0.001 g, 0.0009 g, 0.0008 g, 0.0007 g, 0.0006g, 0.0005 g, 0.0004 g, 0.0003 g, 0.0002 g, or 0.0001 g.

In some embodiments, the amount of one or more of the compounds of thepresent invention is more than 0.0001 g, 0.0002 g, 0.0003 g, 0.0004 g,0.0005 g, 0.0006 g, 0.0007 g, 0.0008 g, 0.0009 g, 0.001 g, 0.0015 g,0.002 g, 0.0025 g, 0.003 g, 0.0035 g, 0.004 g, 0.0045 g, 0.005 g, 0.0055g, 0.006 g, 0.0065 g, 0.007 g, 0.0075 g, 0.008 g, 0.0085 g, 0.009 g,0.0095 g, 0.01 g, 0.015 g, 0.02 g, 0.025 g, 0.03 g, 0.035 g, 0.04 g,0.045 g, 0.05 g, 0.055 g, 0.06 g, 0.065 g, 0.07 g, 0.075 g, 0.08 g,0.085 g, 0.09 g, 0.095 g, 0.1 g, 0.15 g, 0.2 g, 0.25 g, 0.3 g, 0.35 g,0.4 g, 0.45 g, 0.5 g, 0.55 g, 0.6 g, 0.65 g, 0.7 g, 0.75 g, 0.8 g, 0.85g, 0.9 g, 0.95 g, 1 g, 1.5 g, 2 g, 2.5, 3 g, 3.5, 4 g, 4.5 g, 5 g, 5.5g, 6 g, 6.5 g, 7 g, 7.5 g, 8 g, 8.5 g, 9 g, 9.5 g, or 10 g.

In some embodiments, the amount of one or more of the compounds of thepresent invention is in the range of 0.0001-10 g, 0.0005-9 g, 0.001-8 g,0.005-7 g, 0.01-6 g, 0.05-5 g, 0.1-4 g, 0.5-4 g, or 1-3 g.

The combination treatments according to the invention are effective overa wide dosage range. For example, in the treatment of adult humans,dosages from 0.01 to 1000 mg, from 0.5 to 100 mg, from 1 to 50 mg perday, and from 5 to 40 mg per day are examples of dosages that may beused. The exact dosage will depend upon the agent selected, the route ofadministration, the form in which the compound is administered, thesubject to be treated, the body weight of the subject to be treated, andthe preference and experience of the attending physician.

Pharmaceutical Composition for Oral Administration

In some embodiments, the disclosure provides a pharmaceuticalcomposition for oral administration containing at least one compound ofthe present disclosure and a pharmaceutical excipient suitable for oraladministration. The composition may be in the form of a solid, liquid,gel, semi-liquid, or semi-solid. In some embodiments, the compositionfurther comprises a second agent.

In some embodiments, the invention provides a solid pharmaceuticalcomposition for oral administration containing: (i) a HIF-2α inhibitor;(ii) a second compound which is an immunotherapeutic agent; and (iii) apharmaceutical excipient suitable for oral administration. In someembodiments, each compound or agent is present in a therapeuticallyeffective amount. In other embodiments, one or more compounds or agentsis present in a sub-therapeutic amount, and the compounds or agents actsynergistically to provide a therapeutically effective pharmaceuticalcomposition.

In some embodiments, the invention provides for a pharmaceuticalcomposition comprising a combination of a HIF-2α inhibitor and animmunotherapeutic agent. In some embodiments, the HIF-2α inhibitor andthe immunotherapeutic agent are packaged as a single oral dosage form.In other embodiments, the HIF-2α inhibitor and the immunotherapeuticagent can be packaged as separate dosage forms, such as separatetablets.

Pharmaceutical compositions of the disclosure suitable for oraladministration can be presented as discrete dosage forms, such as hardor soft capsules, cachets, troches, lozenges, or tablets, or liquids oraerosol sprays each containing a predetermined amount of an activeingredient as a powder or in granules, a solution, or a suspension in anaqueous or non-aqueous liquid, an oil-in-water emulsion, or awater-in-oil liquid emulsion, or dispersible powders or granules, orsyrups or elixirs. Such dosage forms can be prepared by any of themethods of pharmacy, which typically include the step of bringing theactive ingredient(s) into association with the carrier. In general, thecomposition are prepared by uniformly and intimately admixing the activeingredient(s) with liquid carriers or finely divided solid carriers orboth, and then, if necessary, shaping the product into the desiredpresentation. For example, a tablet can be prepared by compression ormolding, optionally with one or more accessory ingredients. Compressedtablets can be prepared by compressing in a suitable machine the activeingredient(s) in a free-flowing form such as powder or granules,optionally mixed with an excipient such as, but not limited to, abinder, a lubricant, an inert diluent, and/or a surface active ordispersing agent. Molded tablets can be made by molding in a suitablemachine a mixture of the powdered compound moistened with an inertliquid diluent.

This disclosure further encompasses anhydrous pharmaceutical compositionand dosage forms comprising an active ingredient, since water canfacilitate the degradation of some compounds. For example, water may beadded (e.g., 5%) in the pharmaceutical arts as a means of simulatinglong-term storage in order to determine characteristics such asshelf-life or the stability of formulations over time. Anhydrouspharmaceutical compositions and dosage forms of the disclosure can beprepared using anhydrous or low moisture containing ingredients and lowmoisture or low humidity conditions. Pharmaceutical compositions anddosage forms of the disclosure which contain lactose can be madeanhydrous if substantial contact with moisture and/or humidity duringmanufacturing, packaging, and/or storage is expected. An anhydrouspharmaceutical composition may be prepared and stored such that itsanhydrous nature is maintained. Accordingly, anhydrous compositions maybe packaged using materials known to prevent exposure to water such thatthey can be included in suitable formulary kits. Examples of suitablepackaging include, but are not limited to, hermetically sealed foils,plastic or the like, unit dose containers, blister packs, and strippacks.

An active ingredient can be combined in an intimate admixture with apharmaceutical carrier according to conventional pharmaceuticalcompounding techniques. The carrier can take a wide variety of formsdepending on the form of preparation desired for administration. Inpreparing the composition for an oral dosage form, any of the usualpharmaceutical media can be employed as carriers, such as, for example,water, glycols, oils, alcohols, flavoring agents, preservatives,coloring agents, and the like in the case of oral liquid preparations(such as suspensions, solutions, and elixirs) or aerosols; or carrierssuch as starches, sugars, micro-crystalline cellulose, diluents,granulating agents, lubricants, binders, and disintegrating agents canbe used in the case of oral solid preparations, in some embodimentswithout employing the use of lactose. For example, suitable carriersinclude powders, capsules, and tablets, with the solid oralpreparations. If desired, tablets can be coated by standard aqueous ornonaqueous techniques.

Binders suitable for use in pharmaceutical composition and dosage formsinclude, but are not limited to, corn starch, potato starch, or otherstarches, gelatin, natural and synthetic gums such as acacia, sodiumalginate, alginic acid, other alginates, powdered tragacanth, guar gum,cellulose and its derivatives (e.g., ethyl cellulose, cellulose acetate,carboxymethyl cellulose calcium, sodium carboxymethyl cellulose),polyvinyl pyrrolidone, methyl cellulose, pre-gelatinized starch,hydroxypropyl methyl cellulose, microcrystalline cellulose, and mixturesthereof.

Examples of suitable fillers for use in the pharmaceutical compositionand dosage forms disclosed herein include, but are not limited to, talc,calcium carbonate (e.g., granules or powder), microcrystallinecellulose, powdered cellulose, dextrates, kaolin, mannitol, silicicacid, sorbitol, starch, pre-gelatinized starch, and mixtures thereof.

Disintegrants may be used in the composition of the disclosure toprovide tablets that disintegrate when exposed to an aqueousenvironment. Too much of a disintegrant may produce tablets which maydisintegrate in the bottle. Too little may be insufficient fordisintegration to occur and may alter the rate and extent of release ofthe active ingredient(s) from the dosage form. A sufficient amount ofdisintegrant that is neither too little nor too much to detrimentallyalter the release of the active ingredient(s) may be used to form thedosage forms of the compounds disclosed herein. The amount ofdisintegrant used may vary based upon the type of formulation and modeof administration, and may be readily discernible to those of ordinaryskill in the art. About 0.5 to about 15 weight percent of disintegrant,or about 1 to about 5 weight percent of disintegrant, may be used in thepharmaceutical composition. Disintegrants that can be used to formpharmaceutical composition and dosage forms of the disclosure include,but are not limited to, agar-agar, alginic acid, calcium carbonate,microcrystalline cellulose, croscarmellose sodium, crospovidone,polacrilin potassium, sodium starch glycolate, potato or tapioca starch,other starches, pre-gelatinized starch, other starches, clays, otheralgins, other celluloses, gums or mixtures thereof.

Lubricants which can be used to form pharmaceutical composition anddosage forms of the disclosure include, but are not limited to, calciumstearate, magnesium stearate, mineral oil, light mineral oil, glycerin,sorbitol, mannitol, polyethylene glycol, other glycols, stearic acid,sodium lauryl sulfate, talc, hydrogenated vegetable oil (e.g., peanutoil, cottonseed oil, sunflower oil, sesame oil, olive oil, corn oil, andsoybean oil), zinc stearate, ethyl oleate, ethylaureate, agar, ormixtures thereof. Additional lubricants include, for example, a syloidsilica gel, a coagulated aerosol of synthetic silica, or mixturesthereof. A lubricant can optionally be added, in an amount of less thanabout 1 weight percent of the pharmaceutical composition.

When aqueous suspensions and/or elixirs are desired for oraladministration, the active ingredient therein may be combined withvarious sweetening or flavoring agents, coloring matter or dyes and, ifso desired, emulsifying and/or suspending agents, together with suchdiluents as water, ethanol, propylene glycol, glycerin and variouscombinations thereof.

The tablets can be uncoated or coated by known techniques to delaydisintegration and absorption in the gastrointestinal tract and therebyprovide a sustained action over a longer period. For example, a timedelay material such as glyceryl monostearate or glyceryl distearate canbe employed. Formulations for oral use can also be presented as hardgelatin capsules wherein the active ingredient is mixed with an inertsolid diluent, for example, calcium carbonate, calcium phosphate orkaolin, or as soft gelatin capsules wherein the active ingredient ismixed with water or an oil medium, for example, peanut oil, liquidparaffin or olive oil.

Surfactant which can be used to form pharmaceutical composition anddosage forms of the disclosure include, but are not limited to,hydrophilic surfactants, lipophilic surfactants, and mixtures thereof.That is, a mixture of hydrophilic surfactants may be employed, a mixtureof lipophilic surfactants may be employed, or a mixture of at least onehydrophilic surfactant and at least one lipophilic surfactant may beemployed.

A suitable hydrophilic surfactant may generally have an HLB value of atleast 10, while suitable lipophilic surfactants may generally have anHLB value of or less than about 10. An empirical parameter used tocharacterize the relative hydrophilicity and hydrophobicity of non-ionicamphiphilic compounds is the hydrophilic-lipophilic balance (“HLB”value). Surfactants with lower HLB values are more lipophilic orhydrophobic, and have greater solubility in oils, while surfactants withhigher HLB values are more hydrophilic, and have greater solubility inaqueous solutions. Hydrophilic surfactants are generally considered tobe those compounds having an HLB value greater than about 10, as well asanionic, cationic, or zwitterionic compounds for which the HLB scale isnot generally applicable. Similarly, lipophilic (i.e., hydrophobic)surfactants are compounds having an HLB value equal to or less thanabout 10. However, HLB value of a surfactant is merely a rough guidegenerally used to enable formulation of industrial, pharmaceutical andcosmetic emulsions.

Hydrophilic surfactants may be either ionic or non-ionic. Suitable ionicsurfactants include, but are not limited to, alkylammonium salts;fusidic acid salts; fatty acid derivatives of amino acids,oligopeptides, and polypeptides; glyceride derivatives of amino acids,oligopeptides, and polypeptides; lecithins and hydrogenated lecithins;lysolecithins and hydrogenated lysolecithins; phospholipids andderivatives thereof; lysophospholipids and derivatives thereof;carnitine fatty acid ester salts; salts of alkylsulfates; fatty acidsalts; sodium docusate; acylactylates; mono- and di-acetylated tartaricacid esters of mono- and di-glycerides; succinylated mono- anddi-glycerides; citric acid esters of mono- and di-glycerides; andmixtures thereof.

Within the aforementioned group, ionic surfactants include, by way ofexample: lecithins, lysolecithin, phospholipids, lysophospholipids andderivatives thereof; carnitine fatty acid ester salts; salts ofalkylsulfates; fatty acid salts; sodium docusate; acylactylates; mono-and di-acetylated tartaric acid esters of mono- and di-glycerides;succinylated mono- and di-glycerides; citric acid esters of mono- anddi-glycerides; and mixtures thereof.

Ionic surfactants may be the ionized forms of lecithin, lysolecithin,phosphatidylcholine, phosphatidylethanolamine, phosphatidylglycerol,phosphatidic acid, phosphatidylserine, lysophosphatidylcholine,lysophosphatidylethanolamine, lysophosphatidylglycerol, lysophosphatidicacid, lysophosphatidylserine, PEG-phosphatidylethanolamine,PVP-phosphatidylethanolamine, lactylic esters of fatty acids,stearoyl-2-lactylate, stearoyl lactylate, succinylated monoglycerides,mono/diacetylated tartaric acid esters of mono/diglycerides, citric acidesters of mono/diglycerides, cholylsarcosine, caproate, caprylate,caprate, laurate, myristate, palmitate, oleate, ricinoleate, linoleate,linolenate, stearate, lauryl sulfate, teracecyl sulfate, docusate,lauroyl carnitines, palmitoyl carnitines, myristoyl carnitines, andsalts and mixtures thereof.

Hydrophilic non-ionic surfactants may include, but not limited to,alkylglucosides; alkylmaltosides; alkylthioglucosides; laurylmacrogolglycerides; polyoxyalkylene alkyl ethers such as polyethyleneglycol alkyl ethers; polyoxyalkylene alkylphenols such as polyethyleneglycol alkyl phenols; polyoxyalkylene alkyl phenol fatty acid esterssuch as polyethylene glycol fatty acids monoesters and polyethyleneglycol fatty acids diesters; polyethylene glycol glycerol fatty acidesters; polyglycerol fatty acid esters; polyoxyalkylene sorbitan fattyacid esters such as polyethylene glycol sorbitan fatty acid esters;hydrophilic transesterification products of a polyol with at least onemember of the group consisting of glycerides, vegetable oils,hydrogenated vegetable oils, fatty acids, and sterols; polyoxyethylenesterols, derivatives, and analogues thereof; polyoxyethylated vitaminsand derivatives thereof; polyoxyethylene-polyoxypropylene blockcopolymers; and mixtures thereof; polyethylene glycol sorbitan fattyacid esters and hydrophilic transesterification products of a polyolwith at least one member of the group consisting of triglycerides,vegetable oils, and hydrogenated vegetable oils. The polyol may beglycerol, ethylene glycol, polyethylene glycol, sorbitol, propyleneglycol, pentaerythritol, or a saccharide.

Other hydrophilic-non-ionic surfactants include, without limitation,PEG-10 laurate, PEG-12 laurate, PEG-20 laurate, PEG-32 laurate, PEG-32dilaurate, PEG-12 oleate, PEG-15 oleate, PEG-20 oleate, PEG-20 dioleate,PEG-32 oleate, PEG-200 oleate, PEG-400 oleate, PEG-15 stearate, PEG-32distearate, PEG-40 stearate, PEG-100 stearate, PEG-20 dilaurate, PEG-25glyceryl trioleate, PEG-32 dioleate, PEG-20 glyceryl laurate, PEG-30glyceryl laurate, PEG-20 glyceryl stearate, PEG-20 glyceryl oleate,PEG-30 glyceryl oleate, PEG-30 glyceryl laurate, PEG-40 glyceryllaurate, PEG-40 palm kernel oil, PEG-50 hydrogenated castor oil, PEG-40castor oil, PEG-35 castor oil, PEG-60 castor oil, PEG-40 hydrogenatedcastor oil, PEG-60 hydrogenated castor oil, PEG-60 corn oil, PEG-6caprate/caprylate glycerides, PEG-8 caprate/caprylate glycerides,polyglyceryl-10 laurate, PEG-30 cholesterol, PEG-25 phyto sterol, PEG-30soya sterol, PEG-20 trioleate, PEG-40 sorbitan oleate, PEG-80 sorbitanlaurate, polysorbate 20, polysorbate 80, POE-9 lauryl ether, POE-23lauryl ether, POE-10 oleyl ether, POE-20 oleyl ether, POE-20 stearylether, tocopheryl PEG-100 succinate, PEG-24 cholesterol, polyglyceryl-10oleate, Tween 40, Tween 60, sucrose monostearate, sucrose monolaurate,sucrose monopalmitate, PEG 10-100 nonyl phenol series, PEG 15-100 octylphenol series, and poloxamers.

Suitable lipophilic surfactants include, by way of example only: fattyalcohols; glycerol fatty acid esters; acetylated glycerol fatty acidesters; lower alcohol fatty acids esters; propylene glycol fatty acidesters; sorbitan fatty acid esters; polyethylene glycol sorbitan fattyacid esters; sterols and sterol derivatives; polyoxyethylated sterolsand sterol derivatives; polyethylene glycol alkyl ethers; sugar esters;sugar ethers; lactic acid derivatives of mono- and di-glycerides;hydrophobic transesterification products of a polyol with at least onemember of the group consisting of glycerides, vegetable oils,hydrogenated vegetable oils, fatty acids and sterols; oil-solublevitamins/vitamin derivatives; and mixtures thereof. Within this group,preferred lipophilic surfactants include glycerol fatty acid esters,propylene glycol fatty acid esters, and mixtures thereof, or arehydrophobic transesterification products of a polyol with at least onemember of the group consisting of vegetable oils, hydrogenated vegetableoils, and triglycerides.

In one embodiment, the composition may include a solubilizer to ensuregood solubilization and/or dissolution of the compound of the presentdisclosure and to minimize precipitation of the compound of the presentdisclosure. This can be especially important for composition fornon-oral use, e.g., composition for injection. A solubilizer may also beadded to increase the solubility of the hydrophilic drug and/or othercomponents, such as surfactants, or to maintain the composition as astable or homogeneous solution or dispersion.

Examples of suitable solubilizers include, but are not limited to, thefollowing: alcohols and polyols, such as ethanol, isopropanol, butanol,benzyl alcohol, ethylene glycol, propylene glycol, butanediols andisomers thereof, glycerol, pentaerythritol, sorbitol, mannitol,transcutol, dimethyl isosorbide, polyethylene glycol, polypropyleneglycol, polyvinylalcohol, hydroxypropyl methylcellulose and othercellulose derivatives, cyclodextrins and cyclodextrin derivatives;ethers of polyethylene glycols having an average molecular weight ofabout 200 to about 6000, such as tetrahydrofurfuryl alcohol PEG ether(glycofurol) or methoxy PEG; amides and other nitrogen-containingcompounds such as 2-pyrrolidone, 2-piperidone, ε-caprolactam,N-alkylpyrrolidone, N-hydroxyalkylpyrrolidone, N-alkylpiperidone,N-alkylcaprolactam, dimethylacetamide and polyvinylpyrrolidone; esterssuch as ethyl propionate, tributylcitrate, acetyl triethylcitrate,acetyl tributyl citrate, triethylcitrate, ethyl oleate, ethyl caprylate,ethyl butyrate, triacetin, propylene glycol monoacetate, propyleneglycol diacetate, ε-caprolactone and isomers thereof, δ-valerolactoneand isomers thereof, β-butyrolactone and isomers thereof; and othersolubilizers known in the art, such as dimethyl acetamide, dimethylisosorbide, N-methyl pyrrolidones, monooctanoin, diethylene glycolmonoethyl ether, and water.

Mixtures of solubilizers may also be used. Examples include, but notlimited to, triacetin, triethylcitrate, ethyl oleate, ethyl caprylate,dimethylacetamide, N-methylpyrrolidone, N-hydroxyethylpyrrolidone,polyvinylpyrrolidone, hydroxypropyl methylcellulose, hydroxypropylcyclodextrins, ethanol, polyethylene glycol 200-100, glycofurol,transcutol, propylene glycol, and dimethyl isosorbide. Particularlypreferred solubilizers include sorbitol, glycerol, triacetin, ethylalcohol, PEG-400, glycofurol and propylene glycol.

The amount of solubilizer that can be included is not particularlylimited. The amount of a given solubilizer may be limited to abioacceptable amount, which may be readily determined by one of skill inthe art. In some circumstances, it may be advantageous to includeamounts of solubilizers far in excess of bioacceptable amounts, forexample to maximize the concentration of the drug, with excesssolubilizer removed prior to providing the composition to a patientusing conventional techniques, such as distillation or evaporation. Ifpresent, the solubilizer can be in a weight ratio of 10%, 25%, 50%,100%, or up to about 200% by weight, based on the combined weight of thedrug, and other excipients. If desired, very small amounts ofsolubilizer may also be used, such as 5%, 2%, 1% or even less.Typically, the solubilizer may be present in an amount of about 1% toabout 100%, more typically about 5% to about 25% by weight.

The composition can further include one or more pharmaceuticallyacceptable additives and excipients. Such additives and excipientsinclude, without limitation, detackifiers, anti-foaming agents,buffering agents, polymers, antioxidants, preservatives, chelatingagents, viscomodulators, tonicifiers, flavorants, colorants, odorants,opacifiers, suspending agents, binders, fillers, plasticizers,lubricants, and mixtures thereof.

In addition, an acid or a base may be incorporated into the compositionto facilitate processing, to enhance stability, or for other reasons.Examples of pharmaceutically acceptable bases include amino acids, aminoacid esters, ammonium hydroxide, potassium hydroxide, sodium hydroxide,sodium hydrogen carbonate, aluminum hydroxide, calcium carbonate,magnesium hydroxide, magnesium aluminum silicate, synthetic aluminumsilicate, synthetic hydrocalcite, magnesium aluminum hydroxide,diisopropylethylamine, ethanolamine, ethylenediamine, triethanolamine,triethylamine, triisopropanolamine, trimethylamine,tris(hydroxymethyl)aminomethane (TRIS) and the like. Also suitable arebases that are salts of a pharmaceutically acceptable acid, such asacetic acid, acrylic acid, adipic acid, alginic acid, alkanesulfonicacid, amino acids, ascorbic acid, benzoic acid, boric acid, butyricacid, carbonic acid, citric acid, fatty acids, formic acid, fumaricacid, gluconic acid, hydroquinosulfonic acid, isoascorbic acid, lacticacid, maleic acid, oxalic acid, para-bromophenylsulfonic acid, propionicacid, p-toluenesulfonic acid, salicylic acid, stearic acid, succinicacid, tannic acid, tartaric acid, thioglycolic acid, toluenesulfonicacid, uric acid, and the like. Salts of polyprotic acids, such as sodiumphosphate, disodium hydrogen phosphate, and sodium dihydrogen phosphatecan also be used. When the base is a salt, the cation can be anyconvenient and pharmaceutically acceptable cation, such as ammonium,alkali metals, alkaline earth metals, and the like. Example may include,but not limited to, sodium, potassium, lithium, magnesium, calcium andammonium.

Suitable acids are pharmaceutically acceptable organic or inorganicacids. Examples of suitable inorganic acids include hydrochloric acid,hydrobromic acid, hydriodic acid, sulfuric acid, nitric acid, boricacid, phosphoric acid, and the like. Examples of suitable organic acidsinclude acetic acid, acrylic acid, adipic acid, alginic acid,alkanesulfonic acids, amino acids, ascorbic acid, benzoic acid, boricacid, butyric acid, carbonic acid, citric acid, fatty acids, formicacid, fumaric acid, gluconic acid, hydroquinosulfonic acid, isoascorbicacid, lactic acid, maleic acid, methanesulfonic acid, oxalic acid,para-bromophenylsulfonic acid, propionic acid, p-toluenesulfonic acid,salicylic acid, stearic acid, succinic acid, tannic acid, tartaric acid,thioglycolic acid, toluenesulfonic acid, uric acid and the like.

Pharmaceutical Composition for Topical (e.g., Transdermal) Delivery.

In some embodiments, the invention provides a pharmaceutical compositionfor transdermal delivery containing at least one compound of the presentinvention and a pharmaceutical excipient suitable for transdermaldelivery. For example a pharmaceutical composition for topical deliveryis provided comprising at least one HIF-2α inhibitor and/or animmunotherapeutic agent. The HIF-2α inhibitor and the immunotherapeuticagent may be formulated separately, and may further include a thirdtherapeutic agent.

Composition of the present disclosure can be formulated intopreparations in solid, semi-solid, or liquid forms suitable for local ortopical administration, such as gels, water soluble jellies, creams,lotions, suspensions, foams, powders, slurries, ointments, solutions,oils, pastes, suppositories, sprays, emulsions, saline solutions,dimethylsulfoxide (DMSO)-based solutions. In general, carriers withhigher densities are capable of providing an area with a prolongedexposure to the active ingredients. In contrast, a solution formulationmay provide more immediate exposure of the active ingredient to thechosen area.

The pharmaceutical composition also may comprise suitable solid or gelphase carriers or excipients, which are compounds that allow increasedpenetration of, or assist in the delivery of, therapeutic moleculesacross the stratum corneum permeability barrier of the skin. There aremany of these penetration-enhancing molecules known to those trained inthe art of topical formulation. Examples of such carriers and excipientsinclude, but are not limited to, humectants (e.g., urea), glycols (e.g.,propylene glycol), alcohols (e.g., ethanol), fatty acids (e.g., oleicacid), surfactants (e.g., isopropyl myristate and sodium laurylsulfate), pyrrolidones, glycerol monolaurate, sulfoxides, terpenes(e.g., menthol), amines, amides, alkanes, alkanols, water, calciumcarbonate, calcium phosphate, various sugars, starches, cellulosederivatives, gelatin, and polymers such as polyethylene glycols.

Formulations for topical administration may include ointments, lotions,creams, gels (e.g., poloxamer gel), drops, suppositories, sprays,liquids and powders. Conventional pharmaceutical carriers, aqueous,powder or oily bases, thickeners and the like may be necessary ordesirable. The disclosed compositions can be administered, for example,in a microfiber, polymer (e.g., collagen), nanosphere, aerosol, lotion,cream, fabric, plastic, tissue engineered scaffold, matrix material,tablet, implanted container, powder, oil, resin, wound dressing, bead,microbead, slow release bead, capsule, injectables, intravenous drips,pump device, silicone implants, or any bio-engineered materials.

Another exemplary formulation for use in the methods of the presentdisclosure employs transdermal delivery devices (“patches”). Suchtransdermal patches may be used to provide continuous or discontinuousinfusion of a compound of the present disclosure in controlled amounts,either with or without another agent.

The construction and use of transdermal patches for the delivery ofpharmaceutical agents is well known in the art. See, e.g., U.S. Pat.Nos. 5,023,252, 4,992,445 and 5,001,139. Such patches may be constructedfor continuous, pulsatile, or on demand delivery of pharmaceuticalagents.

Pharmaceutical Composition for Injection.

In some embodiments, the disclosure provides a pharmaceuticalcomposition for injection containing a compound of the presentdisclosure and a pharmaceutical excipient suitable for injection. Forexample a pharmaceutical composition for injection is providedcomprising a HIF-2α inhibitor and/or an immunotherapeutic agent. TheHIF-2α inhibitor and the immunotherapeutic agent may be formulatedseparately, and may further include a third therapeutic agent.Components and amounts of agents in the composition are as describedherein.

The forms in which the novel composition of the present disclosure maybe incorporated for administration by injection include aqueous or oilsuspensions, or emulsions, with sesame oil, corn oil, cottonseed oil, orpeanut oil, as well as elixirs, mannitol, dextrose, or a sterile aqueoussolution, and similar pharmaceutical vehicles.

Aqueous solutions in saline are also conventionally used for injection.Ethanol, glycerol, propylene glycol, liquid polyethylene glycol, and thelike (and suitable mixtures thereof), cyclodextrin derivatives, andvegetable oils may also be employed. The proper fluidity can bemaintained, for example, by the use of a coating, such as lecithin, forthe maintenance of the required particle size in the case of dispersionand by the use of surfactants. The prevention of the action ofmicroorganisms can be brought about by various antibacterial andantifungal agents, for example, parabens, chlorobutanol, phenol, sorbicacid, thimerosal, and the like.

Sterile injectable solutions are prepared by incorporating the compoundof the present disclosure in the required amount in the appropriatesolvent with various other ingredients as enumerated above, as required,followed by filtered sterilization. Generally, dispersions are preparedby incorporating the various sterilized active ingredients into asterile vehicle which contains the basic dispersion medium and therequired other ingredients from those enumerated above. In the case ofsterile powders for the preparation of sterile injectable solutions,certain desirable methods of preparation are vacuum-drying andfreeze-drying techniques which yield a powder of the active ingredientplus any additional desired ingredient from a previouslysterile-filtered solution thereof.

Pharmaceutical Composition for Inhalation.

Compositions for inhalation or insufflation include solutions andsuspensions in pharmaceutically acceptable, aqueous or organic solvents,or mixtures thereof, and powders. The liquid or solid composition maycontain suitable pharmaceutically acceptable excipients as describedvide supra. Preferably, the compositions are administered by the oral ornasal respiratory route for local or systemic effect. Compositions inpreferably pharmaceutically acceptable solvents may be nebulized by useof inert gases. Nebulized solutions may be inhaled directly from thenebulizing device or the nebulizing device may be attached to a facemask tent, or intermittent positive pressure breathing machine.Solution, suspension, or powder compositions may be administered,preferably orally or nasally, from devices that deliver the formulationin an appropriate manner. For example a pharmaceutical composition fordelivery by inhalation is provided comprising at least one HIF-2αinhibitor and/or an immunotherapeutic agent. Compositions comprising aHIF-2α inhibitor and an immunotherapeutic agent may be formulatedseparately and may further include a third therapeutic agent.

Other pharmaceutical composition. Pharmaceutical compositions may alsobe prepared from compositions described herein and one or morepharmaceutically acceptable excipients suitable for sublingual, buccal,rectal, intraosseous, intraocular, intranasal, epidural, or intraspinaladministration. Preparations for such pharmaceutical compositions arewell-known in the art. See, e.g., See, e.g., Anderson, Philip O.;Knoben, James E.; Troutman, William G, eds., Handbook of Clinical DrugData, Tenth Edition, McGraw-Hill, 2002; Pratt and Taylor, eds.,Principles of Drug Action, Third Edition, Churchill Livingston, N.Y.,1990; Katzung, ed., Basic and Clinical Pharmacology, Ninth Edition,McGraw Hill, 20037ybg; Goodman and Gilman, eds., The PharmacologicalBasis of Therapeutics, Tenth Edition, McGraw Hill, 2001; RemingtonsPharmaceutical Sciences, 20th Ed., Lippincott Williams & Wilkins., 2000;Martindale, The Extra Pharmacopoeia, Thirty-Second Edition (ThePharmaceutical Press, London, 1999); all of which are incorporated byreference herein in their entirety.

The compounds of the present invention can also be administered in theform of liposomes. As is known in the art, liposomes are generallyderived from phospholipids or other lipid substances. Liposomes areformed by mono- or multilamellar hydrated liquid crystals that aredispersed in an aqueous medium. Any non-toxic, physiologicallyacceptable and metabolizable lipid capable of forming liposomes can beused. The present compositions in liposome form can contain, in additionto a compound of the present invention, stabilizers, preservatives,excipients, and the like. The preferred lipids are the phospholipids andphosphatidyl cholines (lecithins), both natural and synthetic. Methodsto form liposomes are known in the art. See, for example, Prescott, Ed.,“Methods in Cell Biology”, Volume XIV, ISBN: 978-0-12-564114-2, AcademicPress, New York, N.W., p. 33 (1976) and Medina, Zhu, and Kairemo,“Targeted liposomal drug delivery in cancer”, Current Pharm. Des. 10:2981-2989, 2004. For additional information regarding drug formulationand administration, see “Remington: The Science and Practice ofPharmacy,” Lippincott Williams & Wilkins, Philadelphia, ISBN-10:0781746736, 21^(st) Edition (2005).

The invention also provides kits. The kits include one or more HIF-2αinhibitor, one or more immunothereaputic agent, and/or other compoundsof the present invention as described herein, in suitable packaging withwritten material that can include instructions for use, discussion ofclinical studies, listing of side effects, and the like. Such kits mayalso include information, such as scientific literature references,package insert materials, clinical trial results, and/or summaries ofthese and the like, which indicate or establish the activities and/oradvantages of the composition, and/or which describe dosing,administration, side effects, drug interactions, or other informationuseful to the health care provider. Such information may be based on theresults of various studies, for example, studies using experimentalanimals involving in vivo models and studies based on human clinicaltrials. The kit may further contain another agent. In some embodiments,the compound of the present invention and the agent are provided asseparate compositions in separate containers within the kit. In someembodiments, the compound of the present invention and the agent areprovided as a single composition within a container in the kit. Suitablepackaging and additional articles for use (e.g., measuring cup forliquid preparations, foil wrapping to minimize exposure to air, and thelike) are known in the art and may be included in the kit. Kitsdescribed herein can be provided, marketed and/or promoted to healthproviders, including physicians, nurses, pharmacists, formularyofficials, and the like. Kits may also, in some embodiments, be marketeddirectly to the consumer.

In some embodiments, the subject is a human in need of treatment forcancer, or a precancerous condition or lesion, wherein the cancer ispreferably renal cell carcinoma. Subjects that can be treated withcombination treatments of the present invention, or pharmaceuticallyacceptable salt, ester, prodrug, solvate, hydrate or derivatives of thetherapeutic agents, according to the methods of this invention include,for example, subjects that have been diagnosed as having prostatecancer, such as a prostate cancer selected from the following: anadenocarcinoma or an adenocarcinoma that has migrated to the bone;kidney cancer; psoriasis; restenosis; atherosclerosis; BPH; breastcancer such as a ductal carcinoma in duct tissue in a mammary gland,medullary carcinomas, colloid carcinomas, tubular carcinomas, andinflammatory breast cancer; ovarian cancer, including epithelial ovariantumors such as adenocarcinoma in the ovary and an adenocarcinoma thathas migrated from the ovary into the abdominal cavity; uterine cancer;cervical cancer such as adenocarcinoma in the cervix epithelialincluding squamous cell carcinoma and adenocarcinomas; pancreatic cancersuch as epitheloid carcinoma in the pancreatic duct tissue and anadenocarcinoma in a pancreatic duct; bladder cancer such as atransitional cell carcinoma in urinary bladder, urothelial carcinomas(transitional cell carcinomas), tumors in the urothelial cells that linethe bladder, squamous cell carcinomas, adenocarcinomas, and small cellcancers; leukemia such as acute myeloid leukemia (AML), acutelymphocytic leukemia, chronic lymphocytic leukemia, chronic myeloidleukemia, hairy cell leukemia, myelodysplasia, myeloproliferativedisorders, acute myelogenous leukemia (AML), chronic myelogenousleukemia (CML), mastocytosis, chronic lymphocytic leukemia (CLL),multiple myeloma (MM), and myelodysplastic syndrome (MDS); bone cancer;skin cancer such as basal cell carcinoma, melanoma, squamous cellcarcinoma and actinic keratosis, which is a skin condition thatsometimes develops into squamous cell carcinoma; eye retinoblastoma;cutaneous or intraocular (eye) melanoma; primary liver cancer (cancerthat begins in the liver); thyroid cancer such as papillary, follicular,medullary and anaplastic; AIDS-related lymphoma such as diffuse largeB-cell lymphoma, B-cell immunoblastic lymphoma and small non-cleavedcell lymphoma; Kaposi's Sarcoma; viral-induced cancers includinghepatitis B virus (HBV), hepatitis C virus (HCV), and hepatocellularcarcinoma; human lymphotrophic virus-type 1 (HTLV-1) and adult T-cellleukemia/lymphoma; and human papilloma virus (HPV) and cervical cancer;central nervous system cancers (CNS) such as primary brain tumor, whichincludes gliomas (astrocytoma, anaplastic astrocytoma, or glioblastomamultiforme), Oligodendroglioma, Ependymoma, Meningioma, Lymphoma,Schwannoma, and Medulloblastoma; peripheral nervous system (PNS) cancerssuch as acoustic neuromas and malignant peripheral nerve sheath tumor(MPNST) including neurofibromas and schwannomas, malignant fibrouscytoma, malignant fibrous histiocytoma, malignant meningioma, malignantmesothelioma, and malignant mixed Müllerian tumor; oral cavity andoropharyngeal cancer such as, hypopharyngeal cancer, laryngeal cancer,nasopharyngeal cancer, and oropharyngeal cancer; stomach cancer such aslymphomas, gastric stromal tumors, and carcinoid tumors; testicularcancer such as germ cell tumors (GCTs), which include seminomas andnonseminomas, and gonadal stromal tumors, which include Leydig celltumors and Sertoli cell tumors; thymus cancer such as to thymomas,thymic carcinomas, Hodgkin disease, non-Hodgkin lymphomas carcinoids orcarcinoid tumors; rectal cancer; colon cancer; and lung cancer such asnon-small cell lung cancer (NSCLC), which is divided into squamous cellcarcinomas, adenocarcinomas, and large cell undifferentiated carcinomas,and small cell lung cancer;

This invention further relates to a method for using the compounds orpharmaceutical composition in combination with other tumor treatmentapproaches, including surgery, ionizing radiation, photodynamic therapy,or implants, e.g., with corticosteroids, hormones, or used asradiosensitizers.

One such approach may be, for example, radiation therapy in inhibitingabnormal cell growth or treating the proliferative disorder in themammal. Techniques for administering radiation therapy are known in theart, and these techniques can be used in the combination therapydescribed herein. The administration of the compound of the invention inthis combination therapy can be determined as described herein.

Radiation therapy can be administered through one of several methods, ora combination of methods, including without limitation external-beamtherapy, internal radiation therapy, implant radiation, stereotacticradiosurgery, systemic radiation therapy, radiotherapy and permanent ortemporary interstitial brachytherapy. The term “brachytherapy,” as usedherein, refers to radiation therapy delivered by a spatially confinedradioactive material inserted into the body at or near a tumor or otherproliferative tissue disease site. The term is intended withoutlimitation to include exposure to radioactive isotopes (e.g., At-211,I-131, I-125, Y-90, Re-186, Re-188, Sm-153, Bi-212, P-32, andradioactive isotopes of Lu). Suitable radiation sources for use as acell conditioner of the present invention include both solids andliquids. By way of non-limiting example, the radiation source can be aradionuclide, such as I-125, I-131, Yb-169, Ir-192 as a solid source,I-125 as a solid source, or other radionuclides that emit photons, betaparticles, gamma radiation, or other therapeutic rays. The radioactivematerial can also be a fluid made from any solution of radionuclide(s),e.g., a solution of I-125 or I-131, or a radioactive fluid can beproduced using a slurry of a suitable fluid containing small particlesof solid radionuclides, such as Au-198, Y-90. Moreover, theradionuclide(s) can be embodied in a gel or radioactive micro spheres.

Without being limited by any theory, the compounds of the presentinvention can render abnormal cells more sensitive to treatment withradiation for purposes of killing and/or inhibiting the growth of suchcells. Accordingly, this invention further relates to a method forsensitizing abnormal cells in a mammal to treatment with radiation whichcomprises administering to the mammal an amount of a HIF-2α inhibitorfollowed by administering an amount of an immunotherapeutic agent of thepresent invention, or a pharmaceutically acceptable salt, ester,prodrug, solvate, hydrate or derivative thereof, which in combinedamounts are effective in sensitizing abnormal cells to treatment withradiation. The amount of the compound, salt, or solvate in this methodcan be determined according to the means for ascertaining effectiveamounts of such compounds described herein.

Further therapeutic agents that can be combined with a subject compoundmay be found in Goodman and Gilman's “The Pharmacological Basis ofTherapeutics” Tenth Edition edited by Hardman, Limbird and Gilman or thePhysician's Desk Reference, both of which are incorporated herein byreference in their entirety.

In some embodiments, the compositions and methods further compriseadministering, separately or simultaneously one or more additionalagents (e.g. 1, 2, 3, 4, 5, or more). Additional agents can includethose useful in wound healing. Non-limiting examples of additionalagents include antibiotics (e.g. Aminoglycosides, Cephalosporins,Chloramphenicol, Clindamycin, Erythromycins, Fluoroquinolones,Macrolides, Azolides, Metronidazole, Penicillin's, Tetracycline's,Trimethoprim-sulfamethoxazole, Vancomycin), steroids (e.g. Andranes(e.g. Testosterone), Cholestanes (e.g. Cholesterol), Cholic acids (e.g.Cholic acid), Corticosteroids (e.g. Dexamethasone), Estraenes (e.g.Estradiol), Pregnanes (e.g. Progesterone), narcotic and non-narcoticanalgesics (e.g. Morphine, Codeine, Heroin, Hydromorphone, Levorphanol,Meperidine, Methadone, Oxydone, Propoxyphene, Fentanyl, Methadone,Naloxone, Buprenorphine, Butorphanol, Nalbuphine, Pentazocine),chemotherapy (e.g. anti-cancer drugs such as but not limited toAltretamine, Asparaginase, Bleomycin, Busulfan, Carboplatin, Carmustine,Chlorambucil, Cisplatin, Cladribine, Cyclophosphamide, Cytarabine,Dacarbazine, Diethyl stilbesterol, Ethinyl estradiol, Etoposide,Floxuridine, Fludarabine, Fluorouracil, Flutamide, Goserelin,Hydroxyurea, Idarubicin, Ifosfamide, Leuprolide, Levamisole, Lomustine,Mechlorethamine, Medroxyprogesterone, Megestrol, Melphalan,Mercaptopurine, Methotrexate, Mitomycin, Mitotane, Mitoxantrone,Paclitaxel, pentastatin, Pipobroman, Plicamycin, Prednisone,Procarbazine, Streptozocin, Tamoxifen, Teniposide, Vinblastine,Vincristine), anti-inflammatory agents (e.g. Alclofenac; AlclometasoneDipropionate; Algestone Acetonide; alpha Amylase; Amcinafal; Amcinafide;Amfenac Sodium; Amiprilose Hydrochloride; Anakinra; Anirolac;Anitrazafen; Apazone; Balsalazide Disodium; Bendazac; Benoxaprofen;Benzydamine Hydrochloride; Bromelains; Broperamole; Budesonide;Carprofen; Cicloprofen; Cintazone; Cliprofen; Clobetasol Propionate;Clobetasone Butyrate; Clopirac; Cloticasone Propionate; CormethasoneAcetate; Cortodoxone; Decanoate; Deflazacort; Delatestryl;Depo-Testosterone; Desonide; Desoximetasone; Dexamethasone Dipropionate;Diclofenac Potassium; Diclofenac Sodium; Diflorasone Diacetate;Diflumidone Sodium; Diflunisal; Difluprednate; Diftalone; DimethylSulfoxide; Drocinonide; Endrysone; Enlimomab; Enolicam Sodium;Epirizole; Etodolac; Etofenamate; Felbinac; Fenamole; Fenbufen;Fenclofenac; Fenclorac; Fendosal; Fenpipalone; Fentiazac; Flazalone;Fluazacort; Flufenamic Acid; Flumizole; Flunisolide Acetate; Flunixin;Flunixin Meglumine; Fluocortin Butyl; Fluorometholone Acetate;Fluquazone; Flurbiprofen; Fluretofen; Fluticasone Propionate;Furaprofen; Furobufen; Halcinonide; Halobetasol Propionate; HalopredoneAcetate; Ibufenac; Ibuprofen; Ibuprofen Aluminum; Ibuprofen Piconol;Ilonidap; Indomethacin; Indomethacin Sodium; Indoprofen; Indoxole;Intrazole; Isoflupredone Acetate; Isoxepac; Isoxicam; Ketoprofen;Lofemizole Hydrochloride; Lomoxicam; Loteprednol Etabonate;Meclofenamate Sodium; Meclofenamic Acid; Meclorisone Dibutyrate;Mefenamic Acid; Mesalamine; Meseclazone; Mesterolone;Methandrostenolone; Methenolone; Methenolone Acetate; MethylprednisoloneSuleptanate; Morniflumate; Nabumetone; Nandrolone; Naproxen; NaproxenSodium; Naproxol; Nimazone; Olsalazine Sodium; Orgotein; Orpanoxin;Oxandrolane; Oxaprozin; Oxyphenbutazone; Oxymetholone; ParanylineHydrochloride; Pentosan Polysulfate Sodium; Phenbutazone SodiumGlycerate; Pirfenidone; Piroxicam; Piroxicam Cinnamate; PiroxicamOlamine; Pirprofen; Prednazate; Prifelone; Prodolic Acid; Proquazone;Proxazole; Proxazole Citrate; Rimexolone; Romazarit; Salcolex;Salnacedin; Salsalate; Sanguinarium Chloride; Seclazone; Sermetacin;Stanozolol; Sudoxicam; Sulindac; Suprofen; Talmetacin; Talniflumate;Talosalate; Tebufelone; Tenidap; Tenidap Sodium; Tenoxicam; Tesicam;Tesimide; Testosterone; Testosterone Blends; Tetrydamine; Tiopinac;Tixocortol Pivalate; Tolmetin; Tolmetin Sodium; Triclonide;Triflumidate; Zidometacin; Zomepirac Sodium), or anti-histaminic agents(e.g. Ethanolamines (like diphenhydrmine carbinoxamine), Ethylenediamine(like tripelennamine pyrilamine), Alkylamine (like chlorpheniramine,dexchlorpheniramine, brompheniramine, triprolidine), otheranti-histamines like astemizole, loratadine, fexofenadine,Bropheniramine, Clemastine, Acetaminophen, Pseudoephedrine,Triprolidine).

EXAMPLES

The following examples are given for the purpose of illustrating variousembodiments of the invention and are not meant to limit the presentinvention in any fashion. The present examples, along with the methodsdescribed herein are presently representative of preferred embodiments,are exemplary, and are not intended as limitations on the scope of theinvention. Changes therein and other uses which are encompassed withinthe spirit of the invention as defined by the scope of the claims willoccur to those skilled in the art.

Example 1: Synthesis of3-[(1S)-7-(difluoromethylsulfonyl)-2,2-difluoro-1-hydroxy-indan-4-yl]oxy-5-fluoro-benzonitrile(Compound 15)

Step A: Preparation of3-((7-((difluoromethyl)sulfonyl)-2,3-dihydrospiro[indene-1,2′-[1,3]dioxolan]-4-yl)oxy)-5-fluorobenzonitrile

A mixture of 3-fluoro-5-hydroxy-benzonitrile (1.33 g, 9.7 mmol),7′-(difluoromethylsulfonyl)-4′-fluoro-spiro[1,3-dioxolane-2,1′-indane](1.0 g, 3.24 mmol), and cesium bicarbonate (1.26 g, 6.5 mmol) in1-methyl-2-pyrrolidone (1.8 mL) was heated under N₂ at 110° C.(microwave) for 1 hour and 5 minutes. The reaction was repeated tentimes. The reaction mixtures were combined, diluted with EtOAc, andwashed twice with 1 N NaOH. The combined aqueous layer was extractedwith EtOAc. The EtOAc extracts were combined and washed with brine,dried over Na₂SO₄, filtered, and concentrated to about 100 mL to give asuspension. The suspension was filtered to give3-((7-((difluoromethyl)sulfonyl)-2,3-dihydrospiro[indene-1,2′-[1,3]dioxolan]-4-yl)oxy)-5-fluorobenzonitrileas an off-white solid (6.25 g). The filtrate was diluted with EtOAc,washed with brine (3×), dried over Na₂SO₄, filtered, and concentrated.The residue was purified by flash column chromatography on silica gelwith EtOAc/hexane (0% to 40%) to give additional3-((7-((difluoromethyl)sulfonyl)-2,2-difluoro-2,3-dihydrospiro[indene-1,2′-[1,3]dioxolan]-4-yl)oxy)-5-fluorobenzonitrile(3.3 g, 69% combined yield) as a white solid. LCMS ESI (+) m/z 426(M+H).

Step B: Preparation of3-((7-((difluoromethyl)sulfonyl)-1-oxo-2,3-dihydro-1H-inden-4-yl)oxy)-5-fluorobenzonitrile

A mixture of3-((7-((difluoromethyl)sulfonyl)-2,3-dihydrospiro[indene-1,2′-[1,3]dioxolan]-4-yl)oxy)-5-fluorobenzonitrile(10.9 g, 25.6 mmol) and PPTS (667 mg, 2.66 mmol) in acetone (100mL)/water (15 mL) was heated at 82° C. for 5 hours and then 75° C.overnight. The reaction mixture was cooled to room temperature,concentrated under reduced pressure, diluted with EtOAc, washed withsaturated aqueous NaHCO₃, dried over MgSO₄, filtered, and concentrated.The residue was filtered and washed with water. The solid obtained wasbriefly dried under vacuum at 50° C. and then triturated withEtOAc/hexane to give3-((7-((difluoromethyl)sulfonyl)-1-oxo-2,3-dihydro-1H-inden-4-yl)oxy)-5-fluorobenzonitrile(8 g). Flash column chromatography of the mother liquor on silica gelwith EtOAc/hexane (0% to 80%) provided additional3-((7-((difluoromethyl)sulfonyl)-1-oxo-2,3-dihydro-1H-inden-4-yl)oxy)-5-fluorobenzonitrile(1.3 g, combined 9.3 g, quant. yield). LCMS ESI (+) m/z 382 (M+H).

Step C: Preparation of (E,Z)-3-((1-(butylimino)-7-((difluoromethyl)sulfonyl)-2,3-dihydro-1H-inden-4-yl)oxy)-5-fluorobenzonitrile

A mixture of3-((7-((difluoromethyl)sulfonyl)-1-oxo-2,3-dihydro-1H-inden-4-yl)oxy)-5-fluorobenzonitrile(1.42 g, 3.72 mmol), butylamine (6.0 mL) and 5 drops of trifluoroaceticacid (˜0.1 mL) in benzene (40 mL) was refluxed overnight with removal ofwater using a Dean-Stark trap. The reaction mixture was concentratedunder reduced pressure, diluted with methyl tert-butyl ether, washedwith saturated aqueous NaHCO₃ and brine, dried over Na₂SO₄, filtered,and concentrated. The residue was used in the next step without furtherpurification.

Step D: Preparation of3-((7-((difluoromethyl)sulfonyl)-2,2-difluoro-1-oxo-2,3-dihydro-1H-inden-4-yl)oxy)-5-fluorobenzonitrile

A mixture of (E,Z)-3-((1-(butylimino)-7-((difluoromethyl)sulfonyl)-2,3-dihydro-1H-inden-4-yl)oxy)-5-fluorobenzonitrile(1.29 g, 3 mmol, crude from step C), Selectfluor® (2.62 g, 7.4 mmol) andsodium sulfate (4 g, 28.2 mmol) under N₂ was heated at 82° C. for 4hours. After cooling to room temperature, concentrated HCl (37%, 3 mL)was added. The mixture was stirred at room temperature for 15 minutesand then concentrated under reduced pressure. The residue was dilutedwith methyl t-butyl ether, washed with half saturated aqueous NaHCO₃ andthen brine, dried over Na₂SO₄, filtered, and triturated withEtOAc/hexane to give3-((7-((difluoromethyl)sulfonyl)-2,2-difluoro-1-oxo-2,3-dihydro-1H-inden-4-yl)oxy)-5-fluorobenzonitrileas an off-white solid (0.5 g). The mother liquor was purified by flashcolumn chromatography with EtOAc/hexane (5% to 40%) to give additional3-((7-((difluoromethyl)sulfonyl)-2,2-difluoro-1-oxo-2,3-dihydro-1H-inden-4-yl)oxy)-5-fluorobenzonitrile(0.13 g, 51% combined yield). LCMS ESI (+) m/z 418 (M+H) and 435(M+NH₄).

Step E: Preparation of(S)-3-((7-((difluoromethyl)sulfonyl)-2,2-difluoro-1-hydroxy-2,3-dihydro-1H-inden-4-yl)oxy)-5-fluorobenzonitrile(Compound 15)

An ice cold solution of RuCl(p-cymene)[(R,R)-Ts-DPEN] (0.6 mg) indichloromethane (0.2 mL) was added by syringe under nitrogen to an icecold solution of3-[7-(difluoromethylsulfonyl)-2,2-difluoro-1-oxo-indan-4-yl]oxy-5-fluoro-benzonitrile(28 mg, 0.07 mmol), triethylamine (18.7 μL, 0.13 mmol) and formic acid(7.6 μL, 0.2 mmol) in dichloromethane (0.5 mL) and then placed in arefrigerator at 4° C. overnight. The reaction mixture was directlypurified on preparative TLC with EtOAc/hexane (40%) to give Compound 15(23.4 mg, 0.06 mmol, 83% yield). The ee was determined to be greaterthan 95% by ¹⁹F NMR analysis of the corresponding Mosher ester. LCMS ESI(+) m/z 420 (M+H); ¹H NMR (400 MHz, CDCl₃): δ 7.94 (d, 1H), 7.33-6.98(m, 4H), 6.44 (t, 1H), 5.51 (d, 1H), 3.61-3.45 (m, 2H).

Example 2: Synthesis of(S)-3-((2,2-Difluoro-1-hydroxy-7-(methylsulfonyl)-2,3-dihydro-1H-inden-4-yl)oxy)-5-fluorobenzonitrile(Compound 163)

Step A: Preparation of4′-(3-bromo-5-fluoro-phenoxy)-7′-methylsulfonyl-spiro[1,3-dioxolane-2,1′-indane]

Cesium hydrogen carbonate (142 mg, 0.73 mmol) was added all at once to4′-fluoro-7′-methylsulfonyl-spiro[1,3-dioxolane-2,1′-indane] (100 mg,0.37 mmol) and 3-bromo-5-fluoro-phenol (105 mg, 0.55 mmol) in1-methyl-2-pyrrolidone (1.5 mL) at room temperature in a microwavereaction vial equipped with a stir bar. The flask was flushed withnitrogen then sealed with a crimp cap. The reaction was heated to 150°C. for 7 hours, cooled to ambient temperature then purified directly onreverse phase silica gel (25+M, 14 CV, 20-100% MeCN/water) affording4′-(3-bromo-5-fluoro-phenoxy)-7′-methylsulfonyl-spiro[1,3-dioxolane-2,1′-indane](118 mg, 0.26 mmol, 72% yield).

Step B: Preparation of3-fluoro-5-(7′-methylsulfonylspiro[1,3-dioxolane-2,1′-indane]-4′-yl)oxy-benzonitrile

Dichloro[1;1′-bis(diphenylphosphino)ferrocene]palladium (II)dichloromethane adduct (784 mg, 0.97 mmol) was quickly added to adegassed mixture of4′-(3-bromo-5-fluoro-phenoxy)-7′-methylsulfonyl-spiro[1,3-dioxolane-2,1′-indane](4.3 g, 9.7 mmol), zinc cyanide (1.14 g, 9.7 mmol) and zinc powder (761mg, 11.6 mmol) in DMF (60 mL) under nitrogen. The reaction mixture wasthen warmed to 110° C. for 2 hours. After cooling, the mixture wasfiltered through a pad of celite. The filtrate was diluted with water(100 mL), extracted with MTBE (5×100 mL), washed with brine (100 mL),dried over MgSO₄, filtered and concentrated in vacuo. The crude productwas purified on silica gel (100 g SNAP, 14 CV, 15-100% EtOAc/hexanes)then purified again on silica gel (25 g Ultra SNAP, 14 CV, 0-20%dichloromethane/EtOAc) affording3-fluoro-5-(7′-methylsulfonylspiro[1,3-dioxolane-2,1′-indane]-4′-yl)oxy-benzonitrile(3.77 g, 9.7 mmol, 100% yield).

Step C: Preparation of3-fluoro-5-(7-methylsulfonyl-1-oxo-indan-4-yl)oxy-benzonitrile

Pyridinium para-toluenesulfonate (354 mg, 1.4 mmol) was added all atonce to a solution of3-fluoro-5-(7′-methylsulfonylspiro[1,3-dioxolane-2,1′-indane]-4′-yl)oxy-benzonitrile(550 mg, 1.4 mmol) in acetone (6 mL)/water (2 mL) at room temperatureand then warmed to reflux until completion. The mixture was concentratedin vacuo then purified on silica gel (10 g SNAP, 14 CV, 20-100%EtOAc/hexane) affording3-fluoro-5-(7-methylsulfonyl-1-oxo-indan-4-yl)oxy-benzonitrile (450 mg,1.3 mmol, 92% yield).

Step D: Preparation of 3-[(E,Z)-1-butylimino-7-methylsulfonyl-indan-4-yl]oxy-5-fluoro-benzonitrile

Butan-1-amine (5.15 mL, 52 mmol) was added to3-fluoro-5-(7-methylsulfonyl-1-oxo-indan-4-yl)oxy-benzonitrile (450 mg,1.3 mmol) and trifluoroacetic acid (19.96 μL, 0.26 mmol) in benzene (10mL) at room temperature then warmed to reflux with the azeotropicremoval of water by a Dean-Stark apparatus. Progress of the reaction wasmonitored by ¹H-NMR. When complete, the reaction was cooled to roomtemperature then concentrated in vacuo. The residue was diluted withwater (10 mL), extracted with MTBE (3×10 mL), washed with brine anddried over Na₂SO₄, filtered and concentrated. Crude 3-[(E,Z)-1-butylimino-7-methylsulfonyl-indan-4-yl]oxy-5-fluoro-benzonitrilewas used immediately without purification in the next step.

Step E: Preparation of3-(2,2-difluoro-7-methylsulfonyl-1-oxo-indan-4-yl)oxy-5-fluoro-benzonitrile

Selectfluor® (1.15 g, 3.25 mmol) was added to crude 3-[(E,Z)-1-butylimino-7-methylsulfonyl-indan-4-yl]oxy-5-fluoro-benzonitrile(520 mg, 1.3 mmol) and sodium sulfate (369 mg, 2.6 mmol) in acetonitrile(10 mL) then warmed to reflux for 6 hours. The reaction was cooled toroom temperature, concentrated HCl (1.0 mL, 12 mmol) was added andstirred for 15 minutes. The mixture was diluted with water (10 mL),extracted with EtOAc (3×10 mL), washed with brine (10 mL), dried overMgSO₄, filtered and concentrated in vacuo. The residue was purified onsilica gel (25 g SNAP, 14 CV, 20-100% EtOAc/hexane) afforded3-(2,2-difluoro-7-methylsulfonyl-1-oxo-indan-4-yl)oxy-5-fluoro-benzonitrile(437 mg, 1.2 mmol, 88% yield).

Step F: Preparation of(S)-3-((2,2-difluoro-1-hydroxy-7-(methylsulfonyl)-2,3-dihydro-1H-inden-4-yl)oxy)-5-fluorobenzonitrile(Compound 163)

An ice cold solution of RuCl(p-cymene)[(R,R)-Ts-DPEN] (40.7 mg, 0.06mmol) in CH₂Cl₂ (30 mL) was added by syringe under nitrogen to an icecold solution of3-(2,2-difluoro-7-methylsulfonyl-1-oxo-indan-4-yl)oxy-5-fluoro-benzonitrile(2.44 g, 6.4 mmol), triethylamine (1.78 mL, 12.8 mmol) and formic acid(724 μL, 19.2 mmol) in CH₂Cl₂ (30 mL). The reaction was placed in arefrigerator at 4° C. for 16 hours. The mixture was concentrated to 10mL then purified directly on silica gel (25 g SNAP ULTRA, 14 CV, 10-50%EtOAc/hexane) affording Compound 163 (2.15 g, 5.6 mmol, 87% yield).Enantiomeric excess (98%) was determined by chiral HPLC. Retention timefor (S)-enantiomer: 1.93 minutes; retention time for (R)-enantiomer:2.32 minutes. LCMS ESI (−) 428 (M+HCO₂ ⁻). ¹HNMR (400 MHz, CDCl₃): δ7.93 (d, 1H), 7.27-7.24 (m, 1H), 7.15-7.14 (m, 1H), 7.07-7.03 (m, 1H),7.00 (d, 1H), 5.63-5.58 (m, 1H), 3.56-3.35 (m, 3H), 3.24 (s, 3H).

Example 3: Synthesis ofN-(3-Chlorophenyl-4,6-t2)-4-nitrobenzo[c][1,2,5]oxadiazol-5-amine(Compound 183)

Step A: Synthesis of 3-chlorobenzen-4,6-t₂-amine:3-Chloro-4,6-diiodoaniline (100 mg) was dissolved in methanol (3 mL) andadded with triethylamine (0.1 mL) and submitted for overnight tritiationusing 50 Ci of tritium gas, at room temperature. Labile tritium wasremoved by dissolving the crude reaction mixture in methanol (3 mL) andbringing to dryness under vacuum. Labile removal was done in duplicate.The crude tritiated material was purified by preparative TLC (Silicagel, 1000μ) using hexane:ethyl acetate:AcOH (85:14:1). The product bandwas eluted with ethyl acetate to give 3-chlorobenzen-4,6-t₂-amine(yield=600 mCi, radiochemical purity was >98%).

Step B: Synthesis of Compound 183: A stirred mixture of5-chloro-4-nitro-2,1,3-benzoxadiazole (20 mg, 0.1 mmol),3-chlorobenzen-4,6-t₂-amine (600 mCi) and Cs₂CO₃ (65 mg, 0.20 mmol) inDMF (1 mL) was heated at 60° C. for 1 h. After cooling, the reactionmixture was partitioned between EtOAc and water. The aqueous layer wasextracted with EtOAc. The combined organic layers were washed with waterand brine, dried and concentrated. The residue was purified bypreparative HPLC on an ACE-5 C18 Semi-prep column, 250×10 mm, 100 Å.Elution was carried out isocratically using 0.1% TFA inwater/Acetonitrile (35:65) to give Compound 183 (478 mCi, 80%).

Example 4: Synthesis of Isomer 1 ofN—((S)-7-(3-Cyano-5-fluorophenoxy)-2,2-difluoro-3-hydroxy-2,3-dihydro-1H-inden-4-yl)(methyl)(oxo)-λ⁶-sulfanylidene)cyanamide(Compound 240)

Step A:N-((7-(3-cyano-5-fluorophenoxy)-3-hydroxy-2,3-dihydro-1H-inden-4-yl)(methyl)(oxo)-λ⁶-sulfanylidene)cyanamide

Sodium hydrogen carbonate (79.3 mg, 0.94 mmol) was added to a solutionof 3-fluoro-5-hydroxy-benzonitrile (86.27 mg, 0.63 mmol) andN-((7-fluoro-3-hydroxy-2,3-dihydro-1H-inden-4-yl)(methyl)(oxo)-λ⁶-sulfanylidene)cyanamide(80 mg, 0.31 mmol) in DMF (3 mL). The vial was sealed and heated at 100°C. over a weekend. The reaction mixture was partitioned between EtOAcand dilute aqueous NaOH. The EtOAc was washed with water, two portionsof brine, dried over MgSO₄, filtered, and evaporated. The residue waschromatographed on a Biotage 25M reverse phase column with a 20% to 90%ACN:water gradient to affordN-((7-(3-cyano-5-fluorophenoxy)-3-hydroxy-2,3-dihydro-1H-inden-4-yl)(methyl)(oxo)-λ⁶-sulfanylidene)cyanamide(80 mg, 0.21 mmol, 69% yield). m/z (ES-API-pos) [M+H]=372.

Step B:N-((7-(3-cyano-5-fluorophenoxy)-3-oxo-2,3-dihydro-1H-inden-4-yl)(methyl)(oxo)-λ⁶-sulfanylidene)cyanamide

Dess-Martin periodinane (192 mg, 0.45 mmol) was added to a solution ofN-((7-(3-cyano-5-fluorophenoxy)-3-hydroxy-2,3-dihydro-1H-inden-4-yl)(methyl)(oxo)-λ⁶-sulfanylidene)cyanamide(200 mg, 0.54 mmol) in dichloromethane (50 mL). After 10 minutes, thereaction mixture was evaporated and the residue was partitioned betweenEtOAc and aqueous sodium thiosulfate and saturated aqueous NaHCO₃. TheEtOAc layer was washed with water, brine, dried over MgSO₄, filtered,and evaporated to affordN-((7-(3-cyano-5-fluorophenoxy)-3-oxo-2,3-dihydro-1H-inden-4-yl)(methyl)(oxo)-λ⁶-sulfanylidene)cyanamide(174 mg, 0.47 mmol, 88% yield) as a colorless film. m/z (ES-API-pos)[M+H]=370.

Step C:(E/Z)-N-((7-(3-cyano-5-fluorophenoxy)-3-((3-methoxypropyl)imino)-2,3-dihydro-1H-inden-4-yl)(methyl)(oxo)-λ⁶-sulfanylidene)cyanamide

Pivalic acid (9.4 mg, 0.09 mmol) was added to a mixture ofN-((7-(3-cyano-5-fluorophenoxy)-3-oxo-2,3-dihydro-1H-inden-4-yl)(methyl)(oxo)-λ⁶-sulfanylidene)cyanamide(170 mg, 0.46 mmol) and 3-methoxypropylamine (0.12 mL, 1.2 mmol) incyclohexane (7 mL) and toluene (7 mL). The mixture was heated at refluxwith a Hickman still attached. After 1 hour, the reaction mixture wasevaporated and the residue was used as is in the next step.

Step D:N-((7-(3-cyano-5-fluorophenoxy)-2,2-difluoro-3-oxo-2,3-dihydro-1H-inden-4-yl)(methyl)(oxo)-λ⁶-sulfanylidene)cyanamide

1-Chloromethyl-4-fluoro-1,4-diazoniabicyclo[2.2.2]octanebis(tetrafluoroborate) (406 mg, 1.15 mmol) was added to a mixture of(E/Z)-N-((7-(3-cyano-5-fluorophenoxy)-3-((3-methoxypropyl)imino)-2,3-dihydro-1H-inden-4-yl)(methyl)(oxo)-λ⁶-sulfanylidene)cyanamide(202 mg, 0.46 mmol) and sodium sulfate (162 mg, 1.15 mmol) inacetonitrile (5 mL). The mixture was heated at 70° C. After 3.5 hours,the reaction mixture was evaporated and the residue was partitionedbetween EtOAc and water. The EtOAc was washed with brine, dried overMgSO₄, filtered, and evaporated. The residue was taken up in EtOAc,absorbed on silica gel, and chromatographed on a Biotage 25 g SNAPcolumn with a 50% to 100% EtOAc:hexane gradient to affordN-((7-(3-cyano-5-fluorophenoxy)-2,2-difluoro-3-oxo-2,3-dihydro-1H-inden-4-yl)(methyl)(oxo)-λ⁶-sulfanylidene)cyanamide(48 mg, 0.118 mmol, 26% yield. m/z (ES-API-pos) [M+H]=406.

Step E:N—(((S)-7-(3-cyano-5-fluorophenoxy)-2,2-difluoro-3-hydroxy-2,3-dihydro-1H-inden-4-yl)(methyl)(oxo)-λ⁶-sulfanylidene)cyanamide(Compound 240)

RuCl(p-cymene)[(R,R)-Ts-DPEN] (1.5 mg, 0.020 mmol) was added to anitrogen-sparged, ice-cold solution ofN-((7-(3-cyano-5-fluorophenoxy)-2,2-difluoro-3-oxo-2,3-dihydro-1H-inden-4-yl)(methyl)(oxo)-λ⁶-sulfanylidene)cyanamide(49 mg, 0.120 mmol), triethylamine (0.022 mL, 0.16 mmol), and formicacid (0.01 mL, 0.24 mmol) in dichloromethane (5 mL). The flask wasplaced in a 4° C. refrigerator over a weekend. The reaction mixture wasevaporated and the residue was chromatographed on a Biotage 10 g SNAPUltra column with a 20% to 80% EtOAc:hexane gradient to afford a solid,which was triturated twice with chloroform to afford Compound 240 (8.6mg, 0.021 mmol, 18% yield) as a single diastereomer in 93% d.e. bychiral chromatography. ¹H NMR (400 MHz, CD₃OD): δ 8.01 (d, 1H),7.54-7.49 (m, 1H), 7.46-7.44 (m, 1H), 7.40-7.36 (m, 1H), 7.20-7.14 (m,1H), 5.56 (d, 1H), 3.78-3.61 (m, 1H), 3.62 (s, 3H), 3.55-3.47 (m, 1H).m/z (ES-API-pos) [M+H]=408.

Example 5: Synthesis of3-[(1S,2S,3R)-2,3-difluoro-1-hydroxy-7-methylsulfonyl-indan-4-yl]oxy-5-fluoro-benzonitrile(Compound 289)

Step A:[(1S,2R)-4-(3-cyano-5-fluoro-phenoxy)-2-fluoro-7-methylsulfonyl-indan-1-yl]acetate

To a stirred solution of3-fluoro-5-[(1S,2R)-2-fluoro-1-hydroxy-7-methylsulfonyl-indan-4-yl]oxy-benzonitrile(2.00 g, 5.47 mmol) in DCM (27 mL) was added 4-(dimethylamino)pyridine(0.2 g, 1.64 mmol) and triethylamine (1.53 mL, 10.9 mmol). Aceticanhydride (1.00 mL, 10.9 mmol) was added dropwise at 0° C. undernitrogen. The reaction mixture was stirred at ambient temperatureovernight. The reaction mixture was diluted with DCM, washed withsaturated aqueous NaHCO₃ and brine, dried and concentrated. The residuewas purified by flash chromatography on silica gel (20-40% EtOAc/hexane)to give[(1S,2R)-4-(3-cyano-5-fluoro-phenoxy)-2-fluoro-7-methylsulfonyl-indan-1-yl]acetate (1.95 g, 87%). LCMS ESI (+) m/z 408 (M+H).

Step B:[(1S,2S,3S)-3-bromo-4-(3-cyano-5-fluoro-phenoxy)-2-fluoro-7-methylsulfonyl-indan-1-yl] acetate and[(1S,2S,3R)-3-bromo-4-(3-cyano-5-fluoro-phenoxy)-2-fluoro-7-methylsulfonyl-indan-1-yl]acetate

To a stirred solution of[(1S,2R)-4-(3-cyano-5-fluoro-phenoxy)-2-fluoro-7-methylsulfonyl-indan-1-yl]acetate (1.95 g, 4.79 mmol) in 1,2-dichloroethane (24 mL) was addedN-bromosuccinimide (0.94 g, 5.27 mmol) and 2,2′-azobisisobutyronitrile(8 mg, 0.05 mmol). The reaction mixture was heated at 80° C. for 3hours. After cooling, the reaction mixture was diluted with DCM, washedwith saturated aqueous NaHCO₃ and brine, dried and concentrated. Theresidue was purified by column chromatography on silica gel (20-30%EtOAc/hexane) to give[(1S,2S,3S)-3-bromo-4-(3-cyano-5-fluoro-phenoxy)-2-fluoro-7-methylsulfonyl-indan-1-yl]acetate (1.52 g, 65%). LCMS ESI (+) m/z 486, 488 (M+H). Further elutionwith 30-50% EtOAc/hexane gave the more polar product[(1S,2S,3R)-3-bromo-4-(3-cyano-5-fluoro-phenoxy)-2-fluoro-7-methylsulfonyl-indan-1-yl]acetate (0.583 g, 25%). LCMS ESI (+) m/z 486, 488 (M+H).

Step C:[(1S,2R,3S)-4-(3-cyano-5-fluoro-phenoxy)-2-fluoro-3-hydroxy-7-methylsulfonyl-indan-1-yl]acetate

To a combined mixture of[(1S,2S,3S)-3-bromo-4-(3-cyano-5-fluoro-phenoxy)-2-fluoro-7-methylsulfonyl-indan-1-yl]acetate and[(1S,2S,3R)-3-bromo-4-(3-cyano-5-fluoro-phenoxy)-2-fluoro-7-methylsulfonyl-indan-1-yl]acetate prepared in Step B (2.05 g, 4.22 mmol) were added1,2-dimethoxyethane (28 mL) and water (0.050 mL) followed by silverperchlorate hydrate (1.42 g, 6.32 mmol). The reaction mixture was heatedat 70° C. for 2 hours. After cooling, the reaction mixture was dilutedwith EtOAc and filtered through Celite. The filtrate was washed withwater and brine, dried and concentrated. The residue was purified byflash chromatography on silica gel (20-50%) to give[(1S,2R,3S)-4-(3-cyano-5-fluoro-phenoxy)-2-fluoro-3-hydroxy-7-methylsulfonyl-indan-1-yl]acetate (0.416 g, 23%) as the less polar product. LCMS ESI (+) m/z 441(M+NH₄ ⁺). Further elution with 60% EtOAc/hexane gave[(1S,2R,3R)-4-(3-cyano-5-fluoro-phenoxy)-2-fluoro-3-hydroxy-7-methylsulfonyl-indan-1-yl]acetate (0.58 g, 32%). LCMS ESI (+) m/z 441 (M+NH₄ ⁺).

Step D:[(1S,2S,3R)-4-(3-cyano-5-fluoro-phenoxy)-2,3-difluoro-7-methylsulfonyl-indan-1-yl]acetate

To a stirred solution of[(1S,2R,3S)-4-(3-cyano-5-fluoro-phenoxy)-2-fluoro-3-hydroxy-7-methylsulfonyl-indan-1-yl]acetate (416 mg, 0.98 mmol) in DCM (10 mL) was added(diethylamino)sulfur trifluoride (DAST) (0.26 mL, 2.0 mmol) at −78° C.under nitrogen. The reaction mixture was allowed to warm to 0° C. andstirred for 15 minutes. The reaction was quenched by saturated aqueousNaHCO₃. The mixture was partitioned between EtOAc and water. The aqueouslayer was extracted with EtOAc. The combined organic layers were washedwith brine, dried and concentrated. The residue was purified by flashchromatography on silica gel (20-40% EtOAc/hexane) to give[(1S,2S,3R)-4-(3-cyano-5-fluoro-phenoxy)-2,3-difluoro-7-methylsulfonyl-indan-1-yl]acetate (310 mg, 74%). LCMS ESI (+) m/z 426 (M+H).

Step E:3-[(1S,2S,3R)-2,3-difluoro-1-hydroxy-7-methylsulfonyl-indan-4-yl]oxy-5-fluoro-benzonitrile(Compound 289)

To a stirred solution of[(1S,2S,3R)-4-(3-cyano-5-fluoro-phenoxy)-2,3-difluoro-7-methylsulfonyl-indan-1-yl]acetate (0.23 mmol) in tetrahydrofuran (1.5 mL) was added 0.5 N LiOHsolution (0.68 mL, 0.34 mmol) at 0° C. under nitrogen. The reactionmixture was stirred at 0° C. for 1 hour. The reaction was thenpartitioned between EtOAc and water. The aqueous layer was extractedwith EtOAc. The combined organic layers were washed with water andbrine, dried and concentrated. The residue was purified by flashchromatography on silica gel (30-70% EtOAc/hexane) to give Compound 289.LCMS ESI (+) m/z 384 (M+H); ¹H NMR (400 MHz, CDCl₃): δ 8.13 (d, 1H),7.31-7.25 (m, 1H), 7.23-7.19 (m, 1H), 7.14-7.09 (m, 1H), 7.04 (d, 1H),6.09-5.91 (m, 1H), 5.87-5.80 (m, 1H), 5.25-5.05 (m, 1H), 3.32 (s, 3H),2.95 (d, 1H).

Example 6: Synthesis of3-fluoro-5-[(1S,3R)-2,2,3-trifluoro-1-hydroxy-7-methylsulfonyl-indan-4-yl]oxy-benzonitrile(Compound 292)

Step A:[(1S,3S)-4-(3-cyano-5-fluoro-phenoxy)-2,2-difluoro-3-hydroxy-7-methylsulfonyl-indan-1-yl]acetate and[(1S,3R)-4-(3-cyano-5-fluoro-phenoxy)-2,2-difluoro-3-hydroxy-7-methylsulfonyl-indan-1-yl] acetate

To a stirred solution of[(1S)-4-(3-cyano-5-fluoro-phenoxy)-2,2-difluoro-7-methylsulfonyl-indan-1-yl] acetate (1.0 g, 2.35 mmol) in DCE (24 mL) wereadded N-bromosuccinimide (0.46 g, 2.59 mmol) and2,2′-azobisisobutyronitrile (4 mg, 0.02 mmol). The reaction mixture washeated at 80° C. overnight. After cooling, the reaction mixture wasdiluted with DCM, washed with saturated aqueous NaHCO₃ and brine, driedand concentrated. The crude product was dissolved in 1,2-dimethoxyethane(11 mL) and water (0.11 mL). Silver perchlorate hydrate (0.35 g, 1.55mmol) was added. The reaction mixture was heated at 70° C. overnight.After cooling, the reaction mixture was diluted with EtOAc and filteredthrough Celite. The filtrate was washed with water and brine, dried andconcentrated. The residue was purified by flash chromatography on silicagel (20-60% EtOAc/hexane) to give[(1S,3S)-4-(3-cyano-5-fluoro-phenoxy)-2,2-difluoro-3-hydroxy-7-methylsulfonyl-indan-1-yl]acetate (39 mg, 9% yield) as the less polar product. LCMS ESI (+) m/z459 (M+NH₄ ⁺). Further elution gave[(1S,3R)-4-(3-cyano-5-fluoro-phenoxy)-2,2-difluoro-3-hydroxy-7-methylsulfonyl-indan-1-yl]acetate (80 mg, 18%). LCMS ESI (+) m/z 459 (M+NH₄ ⁺).

Step B:[(1S,3R)-4-(3-cyano-5-fluoro-phenoxy)-2,2,3-trifluoro-7-methylsulfonyl-indan-1-yl]acetate

Prepared as described in Example 5 Step D substituting[(1S,2R,3S)-4-(3-cyano-5-fluoro-phenoxy)-2-fluoro-3-hydroxy-7-methylsulfonyl-indan-1-yl]acetate with[(1S,3S)-4-(3-cyano-5-fluoro-phenoxy)-2,2-difluoro-3-hydroxy-7-methylsulfonyl-indan-1-yl]acetate.LCMS ESI (+) m/z 444 (M+H).

Step C:3-fluoro-5-[(1S,3R)-2,2,3-trifluoro-1-hydroxy-7-methylsulfonyl-indan-4-yl]oxy-benzonitrile(Compound 292)

Prepared as described in Example 5 Step E substituting[(1R)-4-(3-cyano-5-fluoro-phenoxy)-3,3-difluoro-7-methylsulfonyl-indan-1-yl]acetatewith[(1S,3R)-4-(3-cyano-5-fluoro-phenoxy)-2,2,3-trifluoro-7-methylsulfonyl-indan-1-yl]acetate. LCMS ESI (+) m/z 419 (M+NH₄ ⁺); ¹H NMR (400 MHz, CDCl₃): δ8.14-8.11 (m, 1H), 7.33-7.29 (m, 1H), 7.25-7.23 (m, 1H), 7.16-7.12 (m,1H), 7.05 (d, 1H), 5.91-5.75 (m, 1H), 5.71-5.65 (m, 1H), 3.39 (d, 1H),3.25 (s, 3H).

Example 7: Synthesis of(R)-3-((4-(Difluoromethyl)-2,2-difluoro-3-hydroxy-1,1-dioxido-2,3-dihydrobenzo[b]thiophen-5-yl)oxy)-5-fluorobenzonitrile(Compound 349)

Step A: Preparation of 2-bromo-3-(difluoromethyl)-1,4-difluorobenzene: Asolution of 2-bromo-3,6-difluorobenzaldehyde (40.0 g, 181 mmol)dissolved in dichloromethane (800 mL) was cooled to 0° C., then treatedwith (diethylamino)sulfur trifluoride (70.0 g, 454 mmol). After theaddition, the reaction mixture was warmed to ambient temperature andstirred at this temperature for 4 hours. Saturated aqueous sodiumbicarbonate solution was added slowly until the pH was 8-9. The organiclayer was separated, dried over sodium sulfate, filtered andconcentrated under reduced pressure to give2-bromo-3-(difluoromethyl)-1,4-difluorobenzene (44.0 g, quant.) as solidwhich was used immediately in the next step without purification. ¹H NMR(400 MHz, CDCl₃): δ 7.28-7.22 (m, 1H), 7.17-7.10 (m, 1H), 7.04 (t, 1H).

Step B: Preparation of 2-(difluoromethyl)-3,6-difluorobenzonitrile: Asuspension of 2-bromo-3-(difluoromethyl)-1,4-difluorobenzene (44.0 g,181 mmol) and copper (I) cyanide (21.1 g, 235 mmol) in1-methyl-2-pyrrolidinone (400 mL) was heated to 180° C. for 2 hours.After cooling to ambient temperature, the reaction mixture was pouredinto water and extracted with diethyl ether. The organic layer waswashed with brine, dried over sodium sulfate, filtered and thenconcentrated under reduced pressure. The crude product was purified byflash chromatography on silica gel eluting with hexane/ethyl acetate togive 2-(difluoromethyl)-3,6-difluorobenzonitrile as a solid (23 g, 67%).¹H NMR (400 MHz, CDCl₃): δ 7.48-7.35 (m, 2H), 6.98 (t, 1H).

Step C: Preparation of2-(difluoromethyl)-3-fluoro-6-(methylthio)benzonitrile: A solution of2-(difluoromethyl)-3,6-difluorobenzonitrile (31.3 g, 65.5 mmol) inacetonitrile (500 mL) was cooled to −30° C., then treated with sodiummethanethiolate (12.8 g, 174 mmol). After addition of the solid, thereaction mixture was stirred for 7 hours while maintaining thetemperature between −30° C. and −40° C. A mixture of water (200 mL) andmethyl t-butyl ether (500 mL) were added and the reaction mixture waswarmed to ambient temperature. The organic layer was separated, washedwith brine, dried over sodium sulfate, filtered and concentrated underreduced pressure to give2-(difluoromethyl)-3-fluoro-6-methylsulfanyl-benzonitrile as yellowsolid (36.3 g, 150 mmol, 91%). ¹H NMR (400 MHz, CDCl₃): δ 7.47-7.44 (m,1H), 7.36-7.32 (m, 1H), 6.99 (t, 1H), 2.58 (s, 3H).

Step D: Preparation of2-(difluoromethyl)-3-fluoro-6-(methylsulfonyl)benzonitrile: A slurry of2-(difluoromethyl)-3-fluoro-6-methylsulfanyl-benzonitrile (36.3 g, 167mmol) in acetonitrile (350 mL) and water (175 mL) was treated withOxone® (257 g, 418 mmol), then the mixture was heated at 56° C. for 4hours. After cooling to ambient temperature, the remaining solids wereremoved by filtration and washed with dichloromethane (300 mL). Thefiltrate was concentrated in vacuo to remove volatile solvents. Theresulting aqueous solution was extracted with dichloromethane (400 mL).The organic layer was dried over sodium sulfate, filtered andconcentrated under reduced pressure. The resulting solid was suspendedin 4:1 hexane/methyl t-butyl ether (200 mL) and stirred for 10 minutesat ambient temperature. The undissolved solid was collected byfiltration and air-dried to give2-(difluoromethyl)-3-fluoro-6-(methylsulfonyl)benzonitrile (29.9 g,71%). ¹H NMR (400 MHz, CDCl₃): δ 8.41-8.37 (m, 1H), 7.66-7.61 (m, 1H),7.11 (t, 1H), 3.34 (s, 3H).

Step E: Preparation of3-(3-cyano-5-fluorophenoxy)-2-(difluoromethyl)-6-(methylsulfonyl)benzonitrile:A suspension of 2-(difluoromethyl)-3-fluoro-6-(methylsulfonyl)benzonitrile (9.52 g, 38.2 mmol),3-fluoro-5-hydroxy-benzonitrile (5.23 g, 38.2 mmol), and cesiumcarbonate (7.77 g, 40.1 mmol) in N, N-dimethylformamide (76 mL) washeated to 45° C. for 3 hours. Additional cesium carbonate (0.46 g, 1.4mmol) was added and the reaction mixture was heated at 45° C. for threehours, then stirred at ambient temperature for 54 hours. The reactionmixture was vigorously stirred while water (800 mL) was added. Theresulting suspension was stirred for 30 minutes, then the solids werecollected by filtration, washed with water (1.2 L), and dried under highvacuum to give 3-(3-cyano-5-fluorophenoxy)-2-(difluoromethyl)-6-(methylsulfonyl)benzonitrile as a white solid (13.3 g, 96%). LCMS ESI (+) m/z384 (M+NH₄). ¹H NMR (400 MHz, DMSO-d₆): δ 8.22 (d, 1H), 7.86-7.82 (m,1H), 7.72-7.62 (m, 3H), 7.49 (t, 1H), 3.44 (s, 3H).

Step F: Preparation of3-((4-(difluoromethyl)-1,1-dioxido-3-oxo-2,3-dihydrobenzo[b]thiophen-5-yl)oxy)-5-fluorobenzonitrile:A solution of3-(3-cyano-5-fluorophenoxy)-2-(difluoromethyl)-6-(methylsulfonyl)benzonitrile(13.3 g, 36 mmol) was dissolved in tetrahydrofuran (380 mL) and treatedwith sodium hydride (60% in mineral oil, 2.26 g, 56 mmol) in two equalportions at five minute intervals. The resulting suspension was stirredat ambient temperature for 60 minutes. The reaction mixture was quenchedby addition of a mixture of 4:1 methanol/10% aqueous HCl (200 mL) andthe resulting suspension was stirred for 1 hour. The mixture wasconcentrated to remove volatile solvents, then the remaining slurry wasdiluted with additional water (800 mL) and stirred for an additional 30minutes. The solids were recovered by filtration and washed withadditional water and the resulting beige solid was dried under highvacuum in the presence of solid NaOH.3-((4-(Difluoromethyl)-1,1-dioxido-3-oxo-2,3-dihydrobenzo[b]thiophen-5-yl)oxy)-5-fluorobenzonitrilewas obtained as a beige solid (13.3 g, quant.) and was used withoutfurther purification. LCMS ESI (−) m/z 366 (M−H). ¹H NMR (400 MHz,DMSO-d₆): δ 8.35 (d, 1H), 7.79 (d, 1H), 7.76 (t, 1H), 7.76-7.72 (m, 1H),7.56-7.50 (m, 2H), 4.72 (s, 2H).

Step G: Preparation of3-((4-(difluoromethyl)-2,2-difluoro-1,1-dioxido-3-oxo-2,3-dihydrobenzo[b]thiophen-5-yl)oxy)-5-fluorobenzonitrile:A solution of3-((4-(difluoromethyl)-1,1-dioxido-3-oxo-2,3-dihydrobenzo[b]thiophen-5-yl)oxy)-5-fluorobenzonitrile(1.40 g, 3.82 mmol) dissolved in acetonitrile (38 mL) was treated atambient temperature with sodium carbonate (890 mg, 8.4 mmol) followed bySelectfluor® (2.98 g, 8.4 mmol). The reaction mixture was stirred atambient temperature for 90 minutes. The reaction mixture wasconcentrated in vacuo to remove volatile solvents, then the residue wasdiluted with water (100 mL) and extracted three times with ethyl acetate(50 mL portions). The combined organic layers were washed with saturatedNaCl, dried over MgSO₄, filtered and concentrated in vacuo to give3-((4-(difluoromethyl)-2,2-difluoro-1,1-dioxido-3-oxo-2,3-dihydrobenzo[b]thiophen-5-yl)oxy)-5-fluorobenzonitrileas a solid (1.48 g, quant.). ¹H NMR (400 MHz, DMSO-d₆, sample exists ashydrate): δ 8.81 (s, 2H), 8.29 (d, 1H), 7.80-7.76 (m, 1H), 7.74 (t, 1H),7.57-7.50 (m, 3H).

Step H: Preparation of3-((4-(difluoromethyl)-2,2-difluoro-3-hydroxy-1,1-dioxido-2,3-dihydrobenzo[b]thiophen-5-yl)oxy)-5-fluorobenzonitrile:A solution of3-((4-(difluoromethyl)-2,2-difluoro-1,1-dioxido-3-oxo-2,3-dihydrobenzo[b]thiophen-5-yl)oxy)-5-fluorobenzonitrile(1.48 g, 3.67 mmol) in methanol (37 mL) was cooled to 0° C., thentreated with sodium borohydride (139 mg, 3.7 mmol) and stirred for 1hour. The reaction was quenched by addition of water (0.5 mL) andsaturated NH₄Cl (0.25 mL). The reaction mixture was concentrated invacuo to remove volatile solvents, then diluted with 0.5 M NaOH (10 mL).The aqueous was extracted three times with ethyl acetate and thecombined organic layers were washed with saturated NaCl, dried overMgSO₄, filtered and concentrated in vacuo. The crude product waschromatographed on SiO₂ eluting with a gradient of ethyl acetate/hexaneto give3-((4-(difluoromethyl)-2,2-difluoro-3-hydroxy-1,1-dioxido-2,3-dihydrobenzo[b]thiophen-5-yl)oxy)-5-fluorobenzonitrileas a white solid (1.24 g, 83%).

Step I: Preparation of(R)-3-((4-(difluoromethyl)-2,2-difluoro-3-hydroxy-1,1-dioxido-2,3-dihydrobenzo[b]thiophen-5-yl)oxy)-5-fluorobenzonitrile:3-((4-(Difluoromethyl)-2,2-difluoro-3-hydroxy-1,1-dioxido-2,3-dihydrobenzo[b]thiophen-5-yl)oxy)-5-fluorobenzonitrilewas resolved using preparative SFC chromatography under the followingconditions: ChiralPak AS(-H) (2×15 cm) column, 20% ethanol with carbondioxide at 100 bar, 60 mL/min flow rate, injection volume was 0.5 mL ofa 20 mg/mL solution in ethanol, peak detection at 220 nm. Compound 349was recovered as the first peak (1.50 minutes) to elute from the column.LCMS ESI (−) m/z 404 (M−H). ¹H NMR (400 MHz, CDCl₃): δ 7.98 (d, 1H),7.33-7.30 (m, 1H), 7.23 (t, 1H), 7.22-7.18 (m, 2H), 7.10-7.06 (m, 1H),5.69-5.65 (m, 1H), 3.23 (d, 1H).

Example 8: Synthesis of(6R,7S)-4-(3,3-difluorocyclobutoxy)-6-fluoro-1-(trifluoromethyl)-6,7-dihydro-5H-cyclopenta[c]pyridin-7-ol(Compound 465) and(R)-4-(3,3-difluorocyclobutoxy)-1-(trifluoromethyl)-6,7-dihydro-5H-cyclopenta[c]pyridin-7-ol(Compound 466)

Step A: Preparation of4-bromo-1-(trifluoromethyl)-5,6-dihydro-7H-cyclopenta[c]pyridin-7-one

A suspension of 4-bromo-5,6-dihydrocyclopenta[c]pyridin-7-one (1.0 g,4.72 mmol) and bis(((trifluoromethyl)sulfinyl)oxy)zinc (4.69 g, 14.15mmol) in a mixture of dichloromethane (30 mL) and water (15 mL) at 0° C.was treated with tert-butyl hydroperoxide (˜70% in water, 2.58 mL, 18.86mmol, added via pipette using a plastic tip) and stirred overnight.Additional portions of bis(((trifluoromethyl)sulfinyl)oxy)zinc (2.35 g,7.07 mmol) and tert-butyl hydroperoxide (2.58 mL, 18.86 mmol) were addedsequentially to drive the reaction to completion. After stirring for anadditional day, the reaction vessel was placed into a water bath andcarefully quenched by the addition of saturated NaHCO₃. Onceeffervescence ceased, the reaction mixture filtered through a pad ofcelite to remove. The pad of celite was rinsed with additionaldichloromethane. The filtrate was separated and the aqueous portionextracted further with 2×20 mL CH₂Cl₂. The combined organics were rinsedwith 10 mL of brine, dried with MgSO₄, filtered, and concentrated todryness. Purification was achieved by chromatography on silica using30-90% CH₂Cl₂/hexane to afford4-bromo-1-(trifluoromethyl)-5,6-dihydro-7H-cyclopenta[c]pyridin-7-one asan off-white solid (390 mg, 30%). The desired regioisomer elutes first.LCMS ESI (+) (M+H) m/z 280/282.

Step B: Preparation of4-bromo-1-(trifluoromethyl)-5,6-dihydrospiro[cyclopenta[c]pyridine-7,2′-[1,3]dioxolane]and4-bromo-1-(trifluoromethyl)-7-(2-((trimethylsilyl)oxy)ethoxy)-5H-cyclopenta[c]pyridine

Trimethylsilyl trifluoromethanesulfonate (75.9 μL, 0.42 mmol) was addedto a solution of4-bromo-1-(trifluoromethyl)-5,6-dihydrocyclopenta[c]pyridin-7-one (389mg, 1.39 mmol) and trimethyl(2-trimethylsilyloxyethoxy)silane (1.37 mL,5.56 mmol) in dichloromethane (13.6 mL) cooled in an ice bath. Themixture was allowed to slowly warm to ambient temperature. After 5 h, anadditional 1.3 mL of trimethyl(2-trimethylsilyloxyethoxy)silane and 76μL of trimethylsilyl trifluoromethanesulfonate were added. After another16 h, the reaction mixture was treated with triethylamine (770 μL, 5.56mmol), stirred for 10 min, and then concentrated. The residue wastreated with 20 mL EtOAc and 20 mL of water and the layers separated.The aqueous portion was extracted further with 2×20 mL of EtOAc. Thecombined organic extracts were washed with brine, dried over MgSO₄,filtered, and evaporated. Purification was achieved by chromatography onsilica using 5-20% EtOAc/hexane to afford4-bromo-1-(trifluoromethyl)-5,6-dihydrospiro[cyclopenta[c]pyridine-7,2′-[1,3]dioxolane]as a shite solid (262 mg, 58%) and4-bromo-1-(trifluoromethyl)-7-(2-((trimethylsilyl)oxy)ethoxy)-5H-cyclopenta[c]pyridineas a white solid (170 mg, 31%). Data for4-bromo-1-(trifluoromethyl)-5,6-dihydrospiro[cyclopenta[c]pyridine-7,2′-[1,3]dioxolane]:LCMS ESI (+) (M+H) m/z 324/326. Data for4-bromo-1-(trifluoromethyl)-7-(2-((trimethylsilyl)ox)ethoxy)-5H-cyclopenta[c]pyridine: ¹H NMR (400 MHz, CDCl₃): δ8.56 (s, 1H), 5.59 (t, 1H), 4.10 (t, 2H), 3.96 (t, 2H), 3.36 (d, 2H),0.15 (s, 9H).

Step C: Preparation of4-bromo-6-fluoro-1-(trifluoromethyl)-5,6-dihydrospiro[cyclopenta[c]pyridine-7,2′-[1,3]dioxolane]

A solution of2-[[4-bromo-1-(trifluoromethyl)-5H-cyclopenta[c]pyridin-7-yl]oxy]ethoxy-trimethyl-silane(146.6 mg, 0.37 mmol) and sodium sulfate (262.7 mg, 1.85 mmol) inacetonitrile (3.7 mL) was stirred for 10 min and then treated withSelectfluor® (145.2 mg, 0.41 mmol) and stirred at 25° C. for 1 h.Volatiles were removed by concentration under reduced pressure. Thereaction mixture was poured into 30 mL of water and extracted with 3×15mL EtOAc. The combined organics were rinsed with 10 mL of brine, driedwith MgSO₄, filtered, and concentrated to dryness. Purification wasachieved by chromatography on silica using 5-20% EtOAc/hexane to afford4-bromo-6-fluoro-1-(trifluoromethyl)-5,6-dihydrospiro[cyclopenta[c]pyridine-7,2′-[1,3]dioxolane]as a white solid (96.2 mg, 76%). LCMS ESI (+) (M+H) m/z 342/344.

Step D: Preparation of6-fluoro-1-(trifluoromethyl)-5,6-dihydrospiro[cyclopenta[c]pyridine-7,2′-[1,3]dioxolan]-4-oland1-(trifluoromethyl)-5,6-dihydrospiro[cyclopenta[c]pyridine-7,2′-[1,3]dioxolan]-4-ol

A solution of4′-bromo-6′-fluoro-1′-(trifluoromethyl)spiro[1,3-dioxolane-2,7′-5,6-dihydrocyclopenta[c]pyridine](96.2mg, 0.2800 mmol) and2-(di-t-butylphosphino)-3,6-dimethoxy-2′,4′,6′-tri-i-propyl-1,1′-biphenyl(3.4 mg, 0.007 mmol) in 1,4-dioxane (5.0 mL) was sparged with nitrogenfor 3 mins. The reaction mixture was then treated sequentially withpotassium hydroxide (47.3 mg, 0.84 mmol), water (101 μL, 5.62 mmol) and[2-(2-aminophenyl)phenyl]-methylsulfonyloxy-palladium;di-t-butyl-[3,6-dimethoxy-2-(2,4,6-triisopropylphenyl)phenyl]phosphane(6.0 mg, 0.007 mmol) under continuous nitrogen stream. The vessel wassealed and heated to 80 C for 1 h and 30 min. The reaction mixture wasquenched by the addition of acetic acid (64.3 μL, 1.13 mmol). Thereaction mixture was poured into 75 mL of water and extracted with 4×20mL EtOAc. The combined organics were dried with MgSO₄, filtered, andconcentrated to dryness. The product was used without furtherpurification (87 mg). During the reaction, some of thehydrodefluorinated product formed as an impurity. Data for6-fluoro-1-(trifluoromethyl)-5,6-dihydrospiro[cyclopenta[c]pyridine-7,2′-[1,3]dioxolan]-4-ol:LCMS ESI (+) (M+H) m/z 280. Data for1-(trifluoromethyl)-5,6-dihydrospiro[cyclopenta[c]pyridine-7,2′-[1,3]dioxolan]-4-ol:LCMS ESI (+) (M+H) m/z 262.

Step E: Preparation of4-(3,3-difluorocyclobutoxy)-6-fluoro-1-(trifluoromethyl)-5,6-dihydrospiro[cyclopenta[c]pyridine-7,2′-[1,3]dioxolane]and4-(3,3-difluorocyclobutoxy)-1-(trifluoromethyl)-5,6-dihydrospiro[cyclopenta[c]pyridine-7,2′-[1,3]dioxolane]

A solution of impure6′-fluoro-1′-(trifluoromethyl)spiro[1,3-dioxolane-2,7′-5,6-dihydrocyclopenta[c]pyridine]-4′-ol(44.0 mg, 0.16 mmol), polymer supported triphenylphosphine (˜2.06mmol/g, 306.2 mg, 0.63 mmol), and 3,3-difluoro-cyclobutanol (68.1 mg,0.63 mmol) in tetrahydrofuran (3.2 mL) was treated with diisopropylazodicarboxylate (120 μL, 0.61 mmol) and stirred at 60° C. for 2 h. Thereaction mixture was filtered and the filter cake rinsed with 20 mLEtOAc. The filtrate was concentrated and purified by chromatography onsilica using 10-30% EtOAc/hexane to afford a clear solid (39.0 mg, 67%)that was a 2:1 mixture of the fluorinated and hydrodefluorinatedproducts. LCMS ESI (+) (M+H) m/z 370. Data for4-(3,3-difluorocyclobutoxy)-6-fluoro-1-(trifluoromethyl)-5,6-dihydrospiro[cyclopenta[c]pyridine-7,2′-[1,3]dioxolane]:LCMS ESI (+) (M+H) m/z 370. Data for4-(3,3-difluorocyclobutoxy)-1-(trifluoromethyl)-5,6-dihydrospiro[cyclopenta[c]pyridine-7,2′-[1,3]dioxolane]:LCMS ESI (+) (M+H) m/z 352.

Step F: Preparation of4-(3,3-difluorocyclobutoxy)-6-fluoro-1-(trifluoromethyl)-5,6-dihydro-7H-cyclopenta[c]pyridin-7-oneand4-(3,3-difluorocyclobutoxy)-1-(trifluoromethyl)-5,6-dihydro-7H-cyclopenta[c]pyridin-7-one

A solution of impure4′-(3,3-difluorocyclobutoxy)-6′-fluoro-1′-(trifluoromethyl)spiro[1,3-dioxolane-2,7′-5,6-dihydrocyclopenta[c]pyridine](39.0 mg, 0.106 mmol) in dichloromethane (2.0 mL) at 0 C was treatedwith perchloric acid (70% in water, 200 μL) and stirred at 0 C for 3 h.The reaction mixture was quenched by the addition of 5 mL of saturatedaqueous NaHCO₃. The resulting mixture was extracted with 3×15 mL CH₂Cl₂.The combined organics were rinsed with 10 mL of brine, dried with MgSO₄,filtered, and concentrated to dryness. The product was used withoutfurther purification as a 2:1 mixture of fluorinated andhydrodefluorinated ketones. LCMS ESI (+) (M+H) m/z 326. Data for4-(3,3-difluorocyclobutoxy)-6-fluoro-1-(trifluoromethyl)-5,6-dihydro-7H-cyclopenta[c]pyridin-7-one:LCMS ESI (+) (M+H) m/z 326. Data for4-(3,3-difluorocyclobutoxy)-1-(trifluoromethyl)-5,6-dihydro-7H-cyclopenta[c]pyridin-7-one:LCMS ESI (+) (M+H) m/z 308.

Step G: Preparation of(6R,7S)-4-(3,3-difluorocyclobutoxy)-6-fluoro-1-(trifluoromethyl)-6,7-dihydro-5H-cyclopenta[c]pyridin-7-ol(Compound 465) and(R)-4-(3,3-difluorocyclobutoxy)-1-(trifluoromethyl)-6,7-dihydro-5H-cyclopenta[c]pyridin-7-ol(Compound 466)

A solution of impure4-(3,3-difluorocyclobutoxy)-6-fluoro-1-(trifluoromethyl)-5,6-dihydrocyclopenta[c]pyridin-7-one(33.8 mg, 0.10 mmol) in dichloromethane (4.0 mL) was cooled to 0° C. andsparged with nitrogen for 5 min. During this time formic acid (11.8 μL,0.31 mmol) and triethylamine (28.8 μL, 0.21 mmol) were sequentiallyadded. Once sparging was complete, RuCl(p-cymene)[(R,R)-Ts-DPEN] (1.3mg, 0.002 mmol) was added under a continuous stream of nitrogen. Thereaction vessel was sealed and placed into the refrigerator to reactovernight. Volatiles were removed by concentration under reducedpressure. The residue was purified by chromatography on silica using4-18% EtOAc/CH₂Cl₂ to afford(6R,7S)-4-(3,3-difluorocyclobutoxy)-6-fluoro-1-(trifluoromethyl)-6,7-dihydro-5H-cyclopenta[c]pyridin-7-ol(Compound 465) as a clear solid (5.4 mg, 16%) and(R)-4-(3,3-difluorocyclobutoxy)-1-(trifluoromethyl)-6,7-dihydro-5H-cyclopenta[c]pyridin-7-ol(Compound 466) as a clear solid (7.4 mg, 23%). Data for(6R,7S)-4-(3,3-difluorocyclobutoxy)-6-fluoro-1-(trifluoromethyl)-6,7-dihydro-5H-cyclopenta[c]pyridin-7-ol(Compound 465): Retention time HPLC (14 min)=3.59 min; LCMS ESI (+)(M+H) m/z 328; ¹H NMR (400 MHz, CDCl₃): δ 8.04 (s, 1H), 5.46-5.26 (m,2H), 4.89-4.79 (m, 1H), 3.36-3.08 (m, 4H), 2.91-2.74 (m, 2H), 2.60 (dd,1H). Data for(R)-4-(3,3-difluorocyclobutoxy)-1-(trifluoromethyl)-6,7-dihydro-5H-cyclopenta[c]pyridin-7-ol(Compound 466): Retention time HPLC (14 min)=3.95 min; LCMS ESI (+)(M+H) m/z 310; ¹H NMR (400 MHz, CDCl₃): δ 7.98 (s, 1H), 5.59-5.54 (m,1H), 4.88-4.79 (m, 1H), 3.24-3.07 (m, 3H), 2.89 (dd, 1H), 2.89-2.74 (m,2H), 2.44-2.34 (m, 1H), 2.28-2.21 (m, 1H), 2.12-2.09 (m, 1H).

Example 9: Synthesis of3-fluoro-5-(((6R,7S)-6-fluoro-7-hydroxy-1-(trifluoromethyl)-6,7-dihydro-5H-cyclopenta[c]pyridin-4-yl)oxy)benzonitrile(Compound 467)

Step A: Preparation of1-(trifluoromethyl)-5,6-dihydrospiro[cyclopenta[c]pyridine-7,2′-[1,3]dioxolan]-4-ol

A solution of4′-bromo-1′-(trifluoromethyl)spiro[1,3-dioxolane-2,7′-5,6-dihydrocyclopenta[c]pyridine](226.4 mg, 0.70 mmol) and2-(di-t-butylphosphino)-3,6-dimethoxy-2′,4′,6′-tri-i-propyl-1,1′-biphenyl(8.5 mg, 0.017 mmol) in 1,4-dioxane (7.0 mL) was sparged with nitrogenfor 3 mins. The reaction mixture was then treated sequentially withpotassium hydroxide (117.6 mg, 2.10 mmol), water (252 μL, 13.97 mmol)and [2-(2-aminophenyl)phenyl]-methylsulfonyloxy-palladium;ditert-butyl-[3,6-dimethoxy-2-(2,4,6-triisopropylphenyl)phenyl]phosphane(14.9 mg, 0.017 mmol) under continuous nitrogen stream. The vessel wassealed and heated to 80 C for 1 h and 30 min. The reaction mixture wasquenched by the addition of acetic acid (160 μL, 2.79 mmol). Thereaction mixture was poured into 75 mL of water and extracted with 4×20mL EtOAc. The combined organics were dried with MgSO4, filtered, andconcentrated to dryness. The brown solid was used without furtherpurification. LCMS ESI (−) (M−H) m/z 260.

Step B: Preparation of3-fluoro-5-((1-(trifluoromethyl)-5,6-dihydrospiro[cyclopenta[c]pyridine-7,2′-[1,3]dioxolan]-4-yl)oxy)benzonitrile

A suspension of potassium tert-butoxide (28.4 mg, 0.25 mmol) intetrahydrofuran (1.5 mL) at 0 C was treated with1′-(trifluoromethyl)spiro[1,3-dioxolane-2,7′-5,6-dihydrocyclopenta[c]pyridine]-4′-ol(60.0 mg, 0.23 mmol) and stirred at 0 C for 15 min. The resultingmixture was treated with(3-cyano-5-fluoro-phenyl)-(4-methoxyphenyl)iodonium;4-methylbenzenesulfonate (144.8 mg, 0.28 mmol) and heated to 40 C. Thereaction mixture was filtered through a plastic filter cup using EtOActo rinse. Volatiles were removed by concentration under reducedpressure. Purification was achieved by chromatography on silica using10-40% EtOAc/hexane to afford a solid (42 mg, 48%). LCMS ESI (+) (M+H)m/z 381.

Step C: Preparation of3-fluoro-5-((7-oxo-1-(trifluoromethyl)-6,7-dihydro-5H-cyclopenta[c]pyridin-4-yl)oxy)benzonitrile

A solution of3-fluoro-5-[1′-(trifluoromethyl)spiro[1,3-dioxolane-2,7′-5,6-dihydrocyclopenta[c]pyridine]-4′-yl]oxy-benzonitrile(42.0 mg, 0.11 mmol) in dichloromethane (2.0 mL) at 0 C was treated withperchloric acid (70% in water, 240 μL) and stirred at 0 C for 30 min.The reaction mixture was carefully quenched by the addition of 15 mL ofsaturated NaHCO₃ and extracted with 3×15 mL CH₂Cl₂. The combinedorganics were rinsed with 10 mL of brine, dried with MgSO₄, filtered,and concentrated to dryness. The solid residue was used immediately inthe next step without further purification. LCMS ESI (+) (M+H) m/z 337.

Step D: Preparation of3-((7-((tert-butyldimethylsilyl)oxy)-1-(trifluoromethyl)-5H-cyclopenta[c]pyridin-4-yl)oxy)-5-fluorobenzonitrile

A solution of triethylamine (122 μL, 0.88 mmol) and3-fluoro-5-[[7-oxo-1-(trifluoromethyl)-5,6-dihydrocyclopenta[c]pyridin-4-yl]oxy]benzonitrile(37.0 mg, 0.11 mmol) in dichloromethane (2.2 mL) at 0° C. was treatedwith tert-butyldimethylsilyl triflate (152 ul, 0.66 mmol). The ice bathwas removed and the reaction mixture left to stir for 2 h. The reactionmixture was poured into 30 mL of saturated NaHCO₃ and extracted with3×20 mL CH₂Cl₂. The combined organics were rinsed with 10 mL of brine,dried with MgSO₄, filtered, and concentrated to dryness. The product wasused without further purification. LCMS ESI (+) (M+H) m/z 451.

Step E: Preparation of3-fluoro-5-((6-fluoro-7-oxo-1-(trifluoromethyl)-6,7-dihydro-5H-cyclopenta[c]pyridin-4-yl)oxy)benzonitrile

A solution of3-[[7-[tert-butyl(dimethyl)silyl]oxy-1-(trifluoromethyl)-5H-cyclopenta[c]pyridin-4-yl]oxy]-5-fluoro-benzonitrile(49.56 mg, 0.110 mmol) in acetonitrile (2.2 mL) at 25° C. was treatedwith Selectfluor® (42.9 mg, 0.12 mmol) and stirred at 25° C. for 1 h.Volatiles were removed by concentration under reduced pressure Thereaction mixture was poured into 30 mL of water and extracted with 3×10mL EtOAc. The combined organics were rinsed with 10 mL of brine, driedwith MgSO₄, filtered, and concentrated to dryness. Purification wasachieved by chromatography on silica using 10-25% EtOAc/hexane to afforda thin film (37.8 mg, 97%). LCMS ESI (+) (M+H) m/z 355.

Step F: Preparation of3-fluoro-5-(((6R,7S)-6-fluoro-7-hydroxy-1-(trifluoromethyl)-6,7-dihydro-5H-cyclopenta[c]pyridin-4-yl)oxy)benzonitrile(Compound 467)

A solution of3-fluoro-5-[[6-fluoro-7-oxo-1-(trifluoromethyl)-5,6-dihydrocyclopenta[c]pyridin-4-yl]oxy]benzonitrile(15.3 mg, 0.043 mmol) in dichloromethane (1.5 mL) was cooled to 0° C.and sparged with nitrogen for 5 min. During this time formic acid (4.9μL, 0.13 mmol) and triethylamine (12.0 μL, 0.086 mmol) were sequentiallyadded. Once the sparging was complete, RuCl(p-cymene)[(R,R)-Ts-DPEN](0.5 mg, 0.00086 mmol) was added under a continuous stream of nitrogen.The reaction vessel was sealed and placed into the refrigerator to reactovernight. Volatiles were removed by concentration under reducedpressure. The residue was purified by chromatography on silica using10-30% EtOAc/hexane to afford3-fluoro-5-(((6R,7S)-6-fluoro-7-hydroxy-1-(trifluoromethyl)-6,7-dihydro-5H-cyclopenta[c]pyridin-4-yl)oxy)benzonitrile(Compound 467) as a clear solid (11.8 mg, 77%). Retention time HPLC (14min)=4.19 min; LCMS ESI (+) (M+H) m/z 357; ¹H NMR (400 MHz, CDCl₃): δ8.33 (s, 1H), 7.22 (ddd, 1H), 7.10-7.08 (m, 1H), 6.99 (dt, 1H),5.54-5.46 (m, 1H), 5.46-5.28 (m, 1H), 3.26 (ddd, 1H), 3.11 (ddd, 1H),2.67 (dd, 1H).

Example 10: Synthesis of3-fluoro-5-(((6R,7S)-6-fluoro-7-hydroxy-1-(methylsulfonyl)-6,7-dihydro-5H-cyclopenta[c]pyridin-4-yl)oxy)benzonitrile(Compound 468)

Step A: Preparation of 5-bromo-2-(methylthio)nicotinic acid

A solution of 5-bromo-2-fluoro-pyridine-3-carboxylic acid (3.50 g, 15.91mmol) in DMF (62 mL) at 0° C. was treated with potassium carbonate (2.42g, 17.5 mmol) and vigorously stirred at 0° C. for 7 minutes. During thistime, nitrogen was sparged through the solution. Then sodiumthiomethoxide (1.23 g, 16.7 mmol) was added in one portion to thereaction vessel under continuous nitrogen stream. The reaction vesselwas sealed and the ice bath removed. The solution turned from a tancolor to faint yellow. The reaction mixture was left to stir overnight.The reaction mixture was poured onto 10% citric acid solution inducingprecipitation of a white solid. The solid was filtered and rinsedexhaustively with water. Finally, the white solid was dried overnightunder high vacuum in the presence of solid NaOH and used without furtherpurification (3.63 g, 92%). LCMS ESI (+) (M+H) m/z 248/250.

Step B: Preparation of 5-bromo-4-formyl-2-(methylthio)nicotinic acid

A solution of 2,2,6,6-tetramethyl-piperidine (3.26 mL, 19.35 mmol) intetrahydrofuran (40.3 mL) at −50 C was treated with n-butyllithium(˜2.5M in hexanes, 7.09 mL, 17.73 mmol) and stirred for 5 min. Then5-bromo-2-methylsulfanyl-pyridine-3-carboxylic acid (2.00 g, 8.06 mmol)was added via cannula over 30 min as a solution in 60 mL of THF. Theresulting mixture stirred for 30 min at −50 C. The reaction mixture wasquenched by the addition of N,N-dimethylformamide (0.94 mL, 12.09 mmol).15 minutes following addition of the DMF, the reaction mixture wasquenched by the addition of 40 mL of 10% citric acid solution (aqueous)and the reaction warmed to room temperature. After stirring for 30 min,excess THF removed by concentration under reduced pressure. The leftovermixture was poured into 120 mL of 3% citric acid (aqueous) and extractedwith 3×50 mL EtOAc. The combined organics were dried with MgSO₄,filtered, and concentrated to dryness. 2.41 g of an orange solid wasisolated and used without further purification. The material wascontaminated with about 22% citric acid based on proton integration ofthe unpurified NMR spectra. LCMS ESI (+) (M+H) m/z 276/278.

Step C: Preparation of methyl(E)-5-bromo-4-(3-ethoxy-3-oxoprop-1-en-1-yl)-2-(methylthio)nicotinate

A solution of7-bromo-1-hydroxy-4-methylsulfanyl-1H-furo[3,4-c]pyridin-3-one (2.21 g,8.00 mmol), lithium chloride (anhydrous, 339.1 mg, 8.00 mmol) and ethyl2-diethoxyphosphorylacetate (1.60 mL, 8.00 mmol) in acetonitrile (80 mL)at 25 C was treated with 1;8-Diazabicyclo[5.4.0]undec-7-ene (2.87 mL,19.20 mmol) and stirred at 25 C for 2 h. Initially, the solution isheterogenous with the pyridine being insoluble. Upon addition of the DBUthe solution becomes homogeneous and darkens in color. After 1 h, thereaction appears to be mostly complete. In addition a precipitate hasformed. Volatiles were removed by concentration under reduced pressure.The product residue was solubilized with 25 mL of DMF and treated withdimethyl sulfate (1.89 mL, 20.00 mmol). After 2 h, the reaction mixturewas poured into 300 mL of water and extracted with 4×40 mL Et₂O. Thecombined organics were rinsed with 20 mL of brine, dried with MgSO₄,filtered, and concentrated to dryness. Purification was achieved bychromatography on silica using 5-20% EtOAc/hexane to afford methyl(E)-5-bromo-4-(3-ethoxy-3-oxoprop-1-en-1-yl)-2-(methylthio)nicotinate asa white solid (1.90 g, 66%). LCMS ESI (+) (M+H) m/z 360/362.

Step D: Preparation of methyl5-bromo-4-(3-ethoxy-3-oxopropyl)-2-(methylthio)nicotinate

A solution of methyl5-bromo-4-[(E)-3-ethoxy-3-oxo-prop-1-enyl]-2-methylsulfanyl-pyridine-3-carboxylate(1.87 g, 5.19 mmol) and cobalt(ii) chloride hexahydrate (123.5 mg, 0.52mmol) in methanol (20.8 mL) at 0° C. was sparged with nitrogen for 3 minand treated with sodium borohydride (98.2 mg, 2.60 mmol) undercontinuous nitrogen stream. The vessel was sealed and the contentsstirred at 0° C. for 10 min. LCMS at this time indicated partialconsumption of the olefin. An additional portion of sodium borohydride(98.2 mg, 2.60 mmol) was added to drive the reaction to completion. Thereaction mixture was quenched by the addition of 30 mL of saturatedaqueous NH₄Cl. Volatiles were removed by concentration under reducedpressure. The reaction mixture was poured into 30 mL of water andextracted with 3×40 mL EtOAc. The combined organics were rinsed with 10mL of brine, dried with MgSO₄, filtered, and concentrated to dryness.Purification was achieved by chromatography on silica using 5-15%EtOAc/hexane to afford the desired product as a solid (1.08 g, 57%).LCMS ESI (+) (M+H) m/z 362/364.

Step E: Preparation of ethyl4-bromo-1-(methylthio)-5,6-dihydro-7H-cyclopenta[c]pyridin-7-one

A solution of methyl5-bromo-4-(3-ethoxy-3-oxo-propyl)-2-methylsulfanyl-pyridine-3-carboxylate(1.08 g, 2.98 mmol) in tetrahydrofuran (29.8 mL) at −78° C. was treatedwith lithium bis(trimethylsilyl)amide (˜1.0 M in THF, 7.16 mL, 7.16mmol) by dropwise addition over 30 minutes. Once the addition wascomplete, LCMS indicated a small amount of starting material remained soan additional 1 mL of lithium bis(trimethylsilyl)amide was added and thereaction allowed to stir for a further 15 minutes. The reaction mixturewas quenched by the addition of 30 mL of saturated aqueous NH₄Cl. THFwas removed by concentration under reduced pressure. The reactionmixture diluted with 60 mL of EtOAc and an additional 30 mL of water. Athick precipitate formed that can be eliminated by the addition of 10%citric acid solution. The reaction mixture was extracted with 3×30 mLEtOAc. The combined organics were rinsed with 10 mL of brine, dried withMgSO₄, filtered, and concentrated to dryness. The intermediate productwas transferred into a microwave tube using 9.1 mL of DMSO. Theresulting mixture was diluted with 900 μL of water. The vessel wassealed and heated to 150 C by microwave irradiation for 40 min. Thereaction mixture was diluted with 120 mL of water to induceprecipitation of the product and vigorously stirred for 30 min. Theprecipitate was collected, dried overnight under high vacuum in thepresence of solid NaOH, and used without further purification. Beigesolid (705 mg, 92%). LCMS ESI (+) (M+H) m/z 258/260.

Step F: Preparation of4-bromo-1-(methylsulfonyl)-5,6-dihydro-7H-cyclopenta[c]pyridin-7-one

A solution of4-bromo-1-methylsulfanyl-5,6-dihydrocyclopenta[c]pyridin-7-one (364.5mg, 1.41 mmol) in methanol (11.3 mL) at 0° C. was treated with asolution of Oxone® (1.91 g, 3.11 mmol) in water (11.3 mL). The reactionwas left to stir for 24 h. Volatiles were removed by concentration underreduced pressure. The reaction mixture was poured into 40 mL of waterand extracted with 3×20 mL EtOAc. The combined organics were rinsed with10 mL of brine, dried with MgSO₄, filtered, and concentrated to dryness.The off white solid was used without further purification. LCMS ESI (+)(M+H) m/z 290/292.

Step G: Preparation of4-bromo-1-(methylsulfonyl)-5,6-dihydrospiro[cyclopenta[c]pyridine-7,2′-[1,3]dioxolane]

Trimethylsilyl trifluoromethanesulfonate (331 μL, 1.83 mmol) was addedto a solution of4-bromo-1-methylsulfonyl-5,6-dihydrocyclopenta[c]pyridin-7-one (354 mg,1.22 mmol) and trimethyl(2-trimethylsilyloxyethoxy)silane (1.20 mL, 4.88mmol) in dichloromethane (14 mL) cooled in an ice bath. The ice bath wasremoved. After 3 h, an additional portion oftrimethyl(2-trimethylsilyloxyethoxy)silane (1.20 mL, 4.88 mmol) wasadded. The reaction was left to stir overnight. After stirring, for therest of the day, the reaction was quenched by the addition oftriethylamine (1.02 mL, 7.32 mmol). The reaction mixture stirred for 10min. Volatiles were removed and the residue suspended into 30 mL ofsaturated aqueous NaHCO₃ and extracted with 3×20 mL EtOAc. The combinedorganics were rinsed with 10 mL of brine, dried with MgSO4, filtered,and concentrated to dryness. Purification was achieved by chromatographyon silica using 15-50% EtOAc/hexane to afford4-bromo-1-(methylsulfonyl)-5,6-dihydrospiro[cyclopenta[c]pyridine-7,2′-[1,3]dioxolane]as a white solid (74.5 mg, 18%). LCMS ESI (+) (M+H) m/z 334/336. Thebulk of the product was isolated as the enol ether (295 mg, 59%).

Step H: Preparation of1-(methylsulfonyl)-5,6-dihydrospiro[cyclopenta[c]pyridine-7,2′-[1,3]dioxolan]-4-ol

A solution of4′-bromo-1′-methylsulfonyl-spiro[1,3-dioxolane-2,7′-5,6-dihydrocyclopenta[c]pyridine](74.5 mg, 0.22 mmol) and2-(di-t-butylphosphino)-3,6-dimethoxy-2′,4′,6′-tri-i-propyl-1,1′-biphenyl(5.4 mg, 0.011 mmol) in 1,4-dioxane (1.5 mL) was sparged with nitrogenfor 3 mins. The reaction mixture was then treated sequentially withpotassium hydroxide (37.5 mg, 0.67 mmol), water (80.3 μL, 4.46 mmol) and[2-(2-aminophenyl)phenyl]-methylsulfonyloxy-palladium;ditert-butyl-[3,6-dimethoxy-2-(2,4,6-triisopropylphenyl)phenyl]phosphane(9.5 mg, 0.011 mmol) under continuous nitrogen stream. The vessel wassealed and heated to 80 C for 1 h and 30 min. The reaction mixture wasquenched by the addition of acetic acid (51.0 μL, 0.89 mmol). Thereaction mixture was poured into 75 mL of water and extracted with 4×20mL EtOAc. The combined organics were dried with MgSO₄, filtered, andconcentrated to dryness. The product was used without furtherpurification (77.9 mg). LCMS ESI (+) (M+H) m/z 272.

Step I: Preparation of3-fluoro-5-((1-(methylsulfonyl)-5,6-dihydrospiro[cyclopenta[c]pyridine-7,2′-[1,3]dioxolan]-4-yl)oxy)benzonitrile

A suspension of potassium carbonate (30.6 mg, 0.22 mmol) in acetonitrile(1.5 mL) at 25 C was treated with1′-methylsulfonylspiro[1,3-dioxolane-2,7′-5,6-dihydrocyclopenta[c]pyridine]-4′-ol(40.0 mg, 0.15 mmol) and stirred at 25 C for 10 min. The resultingmixture was treated with(3-cyano-5-fluoro-phenyl)-(4-methoxyphenyl)iodonium;4-methylbenzenesulfonate (116.2 mg, 0.22 mmol) and heated to 50 C for 3h. Volatiles were removed by concentration under reduced pressure. Thereaction mixture was poured into 20 mL of water and extracted with 3×15mL EtOAc. The combined organics were rinsed with 10 mL of brine, driedwith MgSO₄, filtered, and concentrated to dryness. Purification wasachieved by chromatography on silica using 20-50% EtOAc/hexane to afford(3-fluoro-5-((1-(methylsulfonyl)-5,6-dihydrospiro[cyclopenta[c]pyridine-7,2′-[1,3]dioxolan]-4-yl)oxy)benzonitrileas a solid (25.1 mg, 44%). LCMS ESI (+) (M+H) m/z 391.

Step J: Preparation of3-fluoro-5-((1-(methylsulfonyl)-7-oxo-6,7-dihydro-5H-cyclopenta[c]pyridin-4-yl)oxy)benzonitrile

A solution of4′-(3,3-difluorocyclobutoxy)-6′-fluoro-1′-methylsulfonyl-spiro[1,3-dioxolane-2,7′-5,6-dihydrocyclopenta[c]pyridine](25.1 mg, 0.064 mmol) in dichloromethane (3.0 mL) at 0 C was treatedwith perchloric acid (70% in water, 330 μL) and stirred at 0 C for 2 hand then at room temperature for 30 min. The reaction mixture wascarefully quenched with 5 mL of saturated NaHCO₃/10 mL of water andextracted with 3×15 mL CH₂Cl₂. The combined organics were rinsed with 10mL of brine, dried with MgSO₄, filtered, and concentrated to dryness.The product was used without further purification. LCMS ESI (+) (M+H)m/z 347.

Step K: Preparation of3-((7-((tert-butyldimethylsilyl)oxy)-1-(methylsulfonyl)-5H-cyclopenta[c]pyridin-4-yl)oxy)-5-fluorobenzonitrile

A solution of triethylamine (71.4 μL, 0.51 mmol) and3-fluoro-5-[[7-oxo-1-(trifluoromethyl)-5,6-dihydrocyclopenta[c]pyridin-4-yl]oxy]benzonitrile(22.2 mg, 0.064 mmol) in dichloromethane (3.0 mL) at 0° C. was treatedwith tert-butyldimethylsilyl trifluoromethanesulfonate (58.2 μL, 0.38mmol). The ice bath was removed and the reaction mixture stirred for 2h. The reaction mixture was poured into 30 mL of saturated NaHCO₃ andextracted with 3×20 mL CH₂Cl₂. The combined organics were rinsed with 10mL of brine, dried with MgSO₄, filtered, and concentrated to dryness.The product was used without further purification. LCMS ESI (+) (M+H)m/z 461.

Step L: Preparation of3-fluoro-5-((6-fluoro-1-(methylsulfonyl)-7-oxo-6,7-dihydro-5H-cyclopenta[c]pyridin-4-yl)oxy)benzonitrile

A solution of3-[[7-[tert-butyl(dimethyl)silyl]oxy-1-(trifluoromethyl)-5H-cyclopenta[c]pyridin-4-yl]oxy]-5-fluoro-benzonitrile(29.5 mg, 0.064 mmol) in acetonitrile (2.6 mL) at 25° C. was treatedwith Selectfluor® (24.9 mg, 0.070 mmol) and stirred at 25° C. for 1 h.Volatiles were removed by concentration under reduced pressure. Thereaction mixture was poured into 30 mL of water and extracted with 3×10mL EtOAc. The combined organics were rinsed with 10 mL of brine, driedwith MgSO₄, filtered, and concentrated to dryness. Purification wasachieved by chromatography on silica using 10-25% EtOAc/hexane to afford3-fluoro-5-((6-fluoro-1-(methylsulfonyl)-7-oxo-6,7-dihydro-5H-cyclopenta[c]pyridin-4-yl)oxy)benzonitrileas a thin film (11.1 mg, 48%). LCMS ESI (+) (M+H) m/z 365.

Step M: Preparation of3-fluoro-5-(((6R,7S)-6-fluoro-7-hydroxy-1-(methylsulfonyl)-6,7-dihydro-5H-cyclopenta[c]pyridin-4-yl)oxy)benzonitrile(Compound 468)

A solution of6-fluoro-4-[(5-fluoro-3-pyridyl)oxy]-1-(trifluoromethyl)-5,6-dihydrocyclopenta[c]pyridin-7-one(11.1 mg, 0.031 mmol) in dichloromethane (2.0 mL) was cooled to 0° C.and sparged with nitrogen for 5 min. During this time formic acid (3.4μL, 0.091 mmol) and triethylamine (8.4 μL, 0.061 mmol) were sequentiallyadded. Once the sparging was complete, RuCl(p-cymene)[(R,R)-Ts-DPEN](0.4 mg, 0.0006 mmol) was added under a continuous stream of nitrogen.The reaction vessel was sealed and put into the refrigerator to reactovernight. Volatiles were removed by concentration under reducedpressure. The residue was purified by chromatography on silica using20-60% EtOAc/hexane to afford3-fluoro-5-(((6R,7S)-6-fluoro-7-hydroxy-1-(methylsulfonyl)-6,7-dihydro-5H-cyclopenta[c]pyridin-4-yl)oxy)benzonitrile(Compound 468) as a white solid (7.0 mg, 63%). Retention time HPLC (14min)=3.16 min; LCMS ESI (+) (M+H) m/z 367; ¹H NMR (400 MHz, CDCl₃): δ8.27 (s, 1H), 7.25 (ddd, 1H), 7.14 (m, 1H), 7.03 (dt, 1H), 5.69 (dt,1H), 5.53-5.36 (m, 1H), 4.24 (d, 1H), 3.34 (s, 3H), 3.31-3.24 (m, 1H),3.09 (ddd, 1H).

Example 11: Synthesis of4-bromo-1-(3,5-difluorophenoxy)-6,7-dihydro-5H-cyclopenta[c]pyridine(Compound 469) and4-(difluoromethylsulfonyl)-1-(3,5-difluorophenoxy)-6,7-dihydro-5H-cyclopenta[c]pyridine(Compound 470)

Step A: 4-bromo-1-chloro-6,7-dihydro-5H-cyclopenta[c]pyridine

A mixture of 4-bromo-2,5,6,7-tetrahydrocyclopenta[c]pyridin-1-one (420mg, 1.96 mmol) and POCl₃ (2.20 mL, 23.6 mmol) was heated at reflux for40 hours. After cooling, excess POCl₃ was removed under reducedpressure. The residue was taken up in EtOAc, washed with saturatedaqueous NaHCO₃ and brine, dried and concentrated. The residue waspurified by column chromatography on silica gel (2-10% EtOAc/hexanes) togive 4-bromo-1-chloro-6,7-dihydro-5H-cyclopenta[c]pyridine (207 mg,45%). LCMS ESI (+) m/z 232/234/236 (M+H)⁺.

Step B:4-bromo-1-(3,5-difluorophenoxy)-6,7-dihydro-5H-cyclopenta[c]pyridine(Compound 469)

A mixture of 4-bromo-1-chloro-6,7-dihydro-5H-cyclopenta[c]pyridine (62mg, 0.27 mmol), 3,5-difluorophenol (38 mg, 0.29 mmol), cesium carbonate(130 mg, 0.400 mmol) and NMP (1.8 mL) was heated at 90° C. overnightunder nitrogen. The reaction mixture was heated to 140° C. and stirredovernight again. After cooling, the reaction mixture was partitionedbetween EtOAc and water. The aqueous layer was extracted with EtOAc. Thecombined organic layers were washed with brine, dried and concentrated.The residue was purified by Biotage C18 reverse phase flashchromatography (20-95% acetonitrile/water) to give4-bromo-1-(3,5-difluorophenoxy)-6,7-dihydro-5H-cyclopenta[c]pyridine(Compound 469, 21 mg, 24%) as a pale yellow solid. LCMS ESI (+) m/z326/328 (M+H)⁺.

Step C:S-[1-(3,5-difluorophenoxy)-6,7-dihydro-5H-cyclopenta[c]pyridin-4-yl]ethanethioate

To a vial containing a solution of4-bromo-1-(3,5-difluorophenoxy)-6,7-dihydro-5H-cyclopenta[c]pyridine (20mg, 0.060 mmol) in 1,4-dioxane (0.3 mL) were addedacetylsulfanylpotassium (8.8 mg, 0.080 mmol) and(5-diphenylphosphanyl-9,9-dimethyl-xanthen-4-yl)-diphenyl-phosphane (4.3mg, 0.010 mmol). The mixture was sparged with nitrogen. Thentris(dibenzylideneacetone)dipalladium(0) (3.4 mg, 0.004 mmol) was added,and the vial was sealed and heated at 110° C. for 4 hours. Aftercooling, the reaction mixture was filtered through Celite. The filtratewas concentrated. The residue was purified by Biotage C18 reverse phaseflash chromatography (10-80% acetonitrile/water) to giveS-[1-(3,5-difluorophenoxy)-6,7-dihydro-5H-cyclopenta[c]pyridin-4-yl]ethanethioate (10 mg, 51%). LCMS ESI (+) m/z 322 (M+H)⁺.

Step D:1-(3,5-difluorophenoxy)-6,7-dihydro-5H-cyclopenta[c]pyridine-4-thiol

To a stirred solution ofS-[1-(3,5-difluorophenoxy)-6,7-dihydro-5H-cyclopenta[c]pyridin-4-yl]ethanethioate(10 mg, 0.030 mmol) in MeOH (0.3 mL) was added 1 N LiOH solution (0.050mL, 0.050 mmol). The reaction mixture was stirred at ambient temperaturefor 30 minutes and then concentrated. To the residue was added water.The pH was added to 2-3 using 0.1 N HCl. The mixture was extracted withEtOAc. The combined organic layers were washed with brine, dried andconcentrated. The crude was used in the next step without furtherpurification. LCMS ESI (+) m/z 280 (M+H)⁺.

Step E:4-(difluoromethylsulfanyl)-1-(3,5-difluorophenoxy)-6,7-dihydro-5H-cyclopenta[c]pyridine

To a stirred solution of crude1-(3,5-difluorophenoxy)-6,7-dihydro-5H-cyclopenta[c]pyridine-4-thiol (9mg, 0.03 mmol) in acetonitrile (0.3 mL) was added potassium hydroxide(36 mg, 0.64 mmol) in water (0.3 mL). The reaction mixture was purgedwith nitrogen and then cooled to −78° C. Bromodifluoromethyldiethylphosphonate (17 mg, 0.060 mmol) was added. The resulting mixturewas allowed to warm to ambient temperature and stirred for 3 hours. Thereaction mixture was partitioned between EtOAc and water. The aqueouslayer was extracted with EtOAc. The combined organics were washed withwater and brine, dried over Na₂SO4, filtered, and concentrated todryness. The crude was used in the next step without furtherpurification. LCMS ESI (+) m/z 330 (M+H)⁺.

Step F:4-(difluoromethylsulfonyl)-1-(3,5-difluorophenoxy)-6,7-dihydro-5H-cyclopenta[c]pyridine(Compound 470)

Sodium periodate (18 mg, 0.080 mmol) was added all at once to crude4-(difluoromethylsulfanyl)-1-(3,5-difluorophenoxy)-6,7-dihydro-5H-cyclopenta[c]pyridine(11 mg, 0.030 mmol) and ruthenium(III) chloride (0.2 mg, 0.001 mmol) inacetonitrile (0.2 mL)/CCl₄ (0.2 mL)/water (0.4 mL). The reaction mixturewas stirred at ambient temperature for 3 hours. Solids were removed byfiltration and rinsed with CH₂Cl₂. The organic layer was separated. Theaqueous layer was extracted with CH₂Cl₂. The combined organics werewashed with brine, dried over Na₂SO₄, filtered and concentrated invacuo. The crude product was purified by column chromatography on silicagel (4-12% EtOAc/hexanes) affording Compound 470 (3 mg, 25% overallyield for three steps) as a white solid. LCMS ESI (+) m/z 362 (M+H)⁺. ¹HNMR (400 MHz, CDCl₃): δ 7.49 (s, 1H), 6.80-6.71 (m, 3H), 6.19 (t, 1H),3.36 (t, 2H), 3.06 (t, 2H), 2.31-2.23 (m, 2H).

Example 12: Synthesis of racemic3-fluoro-5-((6R,7S)-6-fluoro-7-hydroxy-1-(trifluoromethyl)-6,7-dihydro-5H-cyclopenta[c]pyridin-4-yl)benzonitrile(Compound 471)

Step A: Preparation of racemic(6R,7S)-4-bromo-6-fluoro-1-(trifluoromethyl)-6,7-dihydro-5H-cyclopenta[c]pyridin-7-ol

A solution of4-bromo-6-fluoro-1-(trifluoromethyl)-5,6-dihydrocyclopenta[c]pyridin-7-one(6.7 mg, 0.022 mmol) in methanol (0.8 mL) at 0° C. was treated withsodium borohydride (0.9 mg, 0.022 mmol) and stirred at 0° C. for 10 min.The reaction mixture was quenched by the addition of 0.2 mL of saturatedNH₄Cl. Volatiles were removed by concentration under reduced pressure.The reaction mixture was poured into 20 mL of water and extracted with3×10 mL EtOAc. The combined organics were rinsed with 10 mL of brine,dried with MgSO₄, filtered, and concentrated to dryness. The product wasused without further purification. LCMS ESI (+) (M+H) m/z 300/302.

Step B: Preparation of racemic3-fluoro-5-((6R,7S)-6-fluoro-7-hydroxy-1-(trifluoromethyl)-6,7-dihydro-5H-cyclopenta[c]pyridin-4-yl)benzonitrile(Compound 471)

A suspension of racemic(6R,7S)-4-bromo-6-fluoro-1-(trifluoromethyl)-6,7-dihydro-5H-cyclopenta[c]pyridin-7-ol(6.7 mg, 0.022 mmol), 3-cyano-5-fluorophenylboronic acid (5.5 mg, 0.033mmol), cesium fluoride (10.5 mg, 0.069 mmol) andbis(di-tert-butyl(4-dimethylaminophenyl)phosphine)dichloropalladium(II)(0.8 mg, 0.0011 mmol) in a mixture 1,4-dioxane (0.8 mL) and water (80μL) was sparged with nitrogen for 3 mins. The vessel was sealed andheated to 80° C. for 1 h. LCMS indicates product formation. The reactionmixture was poured into 60 mL of water and extracted with 3×20 mL EtOAc.The combined organics were rinsed with 10 mL of brine, dried with MgSO₄,filtered, and concentrated to dryness. Purification was achieved bychromatography on silica using 10-30% EtOAc/hexane to afford racemic3-fluoro-5-((6R,7S)-6-fluoro-7-hydroxy-1-(trifluoromethyl)-6,7-dihydro-5H-cyclopenta[c]pyridin-4-yl)benzonitrile(Compound 471) as an orange solid (1.8 mg, 24%). Retention time HPLC (14min)=3.80 min; LCMS ESI (+) (M+H) m/z 341; ¹H NMR (400 MHz, CDCl₃): δ8.65 (s, 1H), 7.53-7.48 (m, 2H), 7.40 (ddd, 1H), 5.56-5.48 (m, 1H), 5.35(ddt, 1H), 3.41 (ddd, 1H), 3.21 (ddd, 1H), 2.65 (dd, 1H).

Example 13: Synthesisof(S)-1-((1S,3R)-2,2,3-trifluoro-1-hydroxy-7-(methylsulfonyl)-2,3-dihydro-1H-inden-4-yl)piperidin-3-ol(Compound 484)

Step A: Preparation of 4,7-difluoro-1H-indene-1,3(2H)-dione

A solution of 3,6 difluorophthalic anhydride (4.25 g, 23.1 mmol),tert-butyl 3-oxobutanoate (4.29 mL, 25.9 mmol) and acetic anhydride(21.0 mL, 221.6 mmol) at 25° C. was treated with triethylamine (11.7 mL,84.3 mmol) and stirred at ambient temperature for 18 hours. The reactionmixture was cooled to 0° C. and treated with 10% hydrochloric acid (65mL, 211 mmol) by dropwise addition. Once the addition was complete, theice bath was removed and the mixture stirred at ambient for 10 minutes.The mixture was then heated to 75° C. for 10 minutes. During this timegas evolution was observed. The suspension slowly broke up to form aclear red mixture. The reaction mixture was poured into 100 mL of waterand extracted with 3×50 mL CH₂Cl₂. The combined organics were dried withMgSO₄, filtered, and concentrated to dryness. The product was usedwithout further purification.

Step B: Preparation of 2,2,4,7-tetrafluoro-1H-indene-1,3(2H)-dione

A solution of the unpurified 4,7-difluoro-1H-indene-1,3(2H)-dione (4.2g, 23.1 mmol) in acetonitrile (100 mL) cooled in a 25° C. water bath wastreated with sodium carbonate (5.38 g, 50.7 mmol). Selectfluor® (17.97g, 50.7 mmol) was added and the reaction mixture was stirred at ambienttemperature for 1 hour. Volatiles were removed under reduced pressureand the residue was poured into 100 mL of 0.1% HCl and extracted with3×50 mL EtOAc. The combined organics were rinsed with 40 mL of brine,dried with MgSO₄, filtered, and concentrated to dryness. The residue waspurified by flash chromatography on silica gel 1:1 hexane/ethyl acetateto give 2,2,4,7-tetrafluoro-1H-indene-1,3(2H)-dione (3.5 g, 70%) as asolid. ¹H NMR (400 MHz, CDCl₃): δ 7.70 (t, 2H).

Step C: Preparation of(S)-2,2,4,7-tetrafluoro-3-hydroxy-2,3-dihydro-1H-inden-1-one

To a solution of 2,2,4,7-tetrafluoro-1H-indene-1,3(2H)-dione (3.48 g,16.0 mmol) in dichloromethane (150 mL) at 0° C. was added formic acid(600 μL, 16.0 mmol) and triethylamine (1.55 mL, 11.2 mmol). Theresulting mixture was sparged with nitrogen for 5 minutes and thenRuCl(p-cymene)[(S,S)-Ts-DPEN] (203.6 mg, 0.32 mmol) was added. Thereaction vessel was sealed and put into a 4° C. refrigerator to standfor 18 hours. The reaction mixture was poured into 40 mL 1 N HCl. TheCH₂Cl₂ layer was separated and the aqueous layer extracted with ethylacetate (2×50 mL). The combined organics were dried with Na₂SO₄,filtered and concentrated under reduced pressure. The residue waspurified by flash chromatography on silica gel using 25% EtOAc/hexane togive (S)-2,2,4,7-tetrafluoro-3-hydroxy-2,3-dihydro-1H-inden-1-one (2.9g, 83%) as an oil. ¹H NMR (400 MHz, CDCl₃): δ 7.51 (ddd, 1H), 7.29-7.23(m, 1H), 5.44 (dd, 1H), 2.79 (dd, 1H).

Step D: Preparation of(S)-2,2,4-trifluoro-3-hydroxy-7-(methylthio)-2,3-dihydro-1H-inden-1-one

A solution of(S)-2,2,4,7-tetrafluoro-3-hydroxy-2,3-dihydro-1H-inden-1-one (966 mg,4.39 mmol) in acetonitrile (40 mL) at 0° C. was sparged with nitrogenfor 5 minutes and treated with sodium thiomethoxide (353.7 mg, 5.05mmol). The ice bath was removed and the reaction mixture was allowed tostir at ambient temperature for 2 hours. The reaction mixture wasevaporated and the residue partitioned between 40 mL of EtOAc and 40 mLof water. The aqueous layer was further extracted with 2×40 mL of EtOAc.The combined organic extracts were washed with brine, dried over MgSO₄,filtered, and evaporated. The residue was chromatographed on silicausing 10-60% EtOAc/hexane to afford(S)-2,2,4-trifluoro-3-hydroxy-7-(methylthio)-2,3-dihydro-1H-inden-1-one(870 mg, 80%) as a yellow solid. LCMS ESI (+) m/z 249 (M+H).

Step E: Preparation of(S)-2,2,4-trifluoro-3-hydroxy-7-(methylsulfonyl)-2,3-dihydro-1H-inden-1-one

(S)-2,2,4-trifluoro-3-hydroxy-7-(methylthio)-2,3-dihydro-1H-inden-1-one(400 mg, 1.6 mmol) was dissolved in MeOH (10 mL) and the reaction wastreated dropwise with a solution of Oxone® (2.18 g, 3.55 mmol) dissolvedin water (10 mL). The mixture was stirred at ambient temperature for 14hours. The reaction mixture was filtered, the solids were washed withethyl acetate and the filtrate was concentrated in vacuo. The aqueousfiltrate was extracted 3×30 mL of EtOAc and then the combined organicswere washed with saturated NaCl, dried over Na₂SO₄ and concentrated invacuo to a yellow solid that was used without further purification (467mg). LCMS ESI (+) m/z 281.1 (M+H).

Step F: Preparation of(R)-2,2,3,4-tetrafluoro-7-(methylsulfonyl)-2,3-dihydro-1H-inden-1-one

(S)-2,2,4-trifluoro-3-hydroxy-7-(methylsulfonyl)-2,3-dihydro-1H-inden-1-one(450 mg, 1.6 mmol) was dissolved in dichloromethane (16 mL), cooled to0° C. and treated dropwise with diethylaminosulfur trifluoride (0.32 mL,2.4 mmol) and the mixture was stirred at 0° C. for 2 hours, then thewhole homogeneous reaction mixture was placed into the refrigeratorovernight. The reaction was treated with additional diethylaminosulfurtrifluoride (0.32 mL, 2.4 mmol) and stirring continued for 6 hours at 0°C. The cold reaction was treated with saturated NaHCO₃ (10 mL) andstirred vigorously for 20 minutes. The mixture was diluted withadditional methylene chloride and the layers were separated. The aqueouswas re-extracted with methylene chloride and the combined organic layerswere dried over Na₂SO₄ and concentrated in vacuo to a yellow solid. Thecrude material was chromatographed on SiO₂ (Biotage SNAP Ultra) andeluted with a gradient of ethyl acetate/hexanes. The desired materialwas concentrated to a pale yellow solid (258 mg). LCMS ESI (+) m/z 283(M+H).

Step G: Preparation of(R)-2,2,3-trifluoro-4-((S)-3-hydroxypiperidin-1-yl)-7-(methylsulfonyl)-2,3-dihydro-1H-inden-1-one

A solution of (3R)-2,2,3,4-tetrafluoro-7-methylsulfonyl-indan-1-one(28.7 mg, 0.10 mmol) and (3S)-3-piperidinol, hydrochloride (14.0 mg,0.10 mmol) in DMF (700 μL) was treated with cesium bicarbonate (59.2 mg,0.31 mmol) and stirred at 35° C. for 3 h. The reaction mixture waspoured into 30 mL of water and extracted with 3×10 mL Et₂O. The combinedorganics were rinsed with 10 mL of brine, dried with MgSO₄, filtered,and concentrated to dryness. The product was used without furtherpurification. LCMS ESI (+) m/z 364 (M+H).

Step H: Preparationof(S)-1-((1S,3R)-2,2,3-trifluoro-1-hydroxy-7-(methylsulfonyl)-2,3-dihydro-1H-inden-4-yl)piperidin-3-ol(Compound 484)

A solution of(3R)-2,2,3-trifluoro-4-[(3S)-3-hydroxy-1-piperidyl]-7-methylsulfonyl-indan-1-one(36.3 mg, 0.10 mmol) in dichloromethane (4 mL) was cooled to 0° C. andsparged with nitrogen for 5 min. During this time formic acid (12.1 μL,0.32 mmol) and triethylamine (27.9 μL, 0.20 mmol) were sequentiallyadded. Once the sparging was complete, RuCl(p-cymene)[(R,R)-Ts-DPEN](1.3 mg, 2 mol %) was added under a continuous stream of nitrogen. Thereaction vessel was sealed and put into the refrigerator to reactovernight. Volatiles were removed by concentration under reducedpressure. The residue was purified by chromatography on silica using45-75% EtOAc/hexane to afford(S)-1-((1S,3R)-2,2,3-trifluoro-1-hydroxy-7-(methylsulfonyl)-2,3-dihydro-1H-inden-4-yl)piperidin-3-ol(Compound 484) as a white solid (23.9 mg, 65%). Retention time HPLC (14min)=2.63 min; LCMS ESI (+) m/z 366 (M+H); ¹H NMR (400 MHz, CDCl₃): δ7.97 (dd, 1H), 7.07 (d, 1H), 5.74 (dd, 1H), 5.55 (dd, 1H), 4.01-3.93 (m,1H), 3.46-3.34 (m, 3H), 3.33 (d, 1H), 3.20-3.13 (m, 1H), 3.18 (s, 3H),2.01-1.91 (m, 2H), 1.89 (d, 1H), 1.82-1.62 (m, 2H).

Example 14: Synthesis of(S)-2,2-difluoro-4-((1R,3R)-3-hydroxycyclohexyl)-7-((trifluoromethyl)sulfonyl)-2,3-dihydro-1H-inden-1-ol(Compound 480) and(1S)-2,2-difluoro-4-(3-hydroxycyclohex-1-en-1-yl)-7-((trifluoromethyl)sulfonyl)-2,3-dihydro-1H-inden-1-ol(Compound 481) and(S)-2,2-difluoro-4-((1R,3S)-3-hydroxycyclohexyl)-7-((trifluoromethyl)sulfonyl)-2,3-dihydro-1H-inden-1-ol(Compound 482) and(S)-2,2-difluoro-4-((1S,3R)-3-hydroxycyclohexyl)-7-(methylsulfonyl)-2,3-dihydro-1H-inden-1-ol(Compound 483)

Step A: Preparation of(S)-3-(2,2-difluoro-1-hydroxy-7-((trifluoromethyl)sulfonyl)-2,3-dihydro-1H-inden-4-yl)cyclohex-2-en-1-one

A suspension of(1S)-4-bromo-2,2-difluoro-7-(trifluoromethylsulfonyl)indan-1-ol (102.0mg, 0.27 mmol),3-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)cyclohex-2-en-1-one (89.2mg, 0.40 mmol), cesium fluoride (126.0 mg, 0.83 mmol) andbis(di-tert-butyl(4-dimethylaminophenyl)phosphine)dichloropalladium(II)(9.5 mg, 0.013 mmol) in 1,4-dioxane (4.5 mL) was sparged with nitrogenfor 3 mins. The vessel was sealed and heated to 80° C. for 1 h. Thereaction mixture was poured into 60 mL of water and extracted with 3×20mL EtOAc. The combined organics were rinsed with 10 mL of brine, driedwith MgSO₄, filtered, and concentrated to dryness. Purification wasachieved by chromatography on silica using 20-50% EtOAc/hexane. LCMS ESI(−) (M−H) m/z 395.

Step B: Preparation of(S)-2,2-difluoro-4-((1R,3R)-3-hydroxycyclohexyl)-7-((trifluoromethyl)sulfonyl)-2,3-dihydro-1H-inden-1-ol(Compound 480) and(1)-2,2-difluoro-4-(3-hydroxycyclohex-1-en-1-yl)-7-((trifluoromethyl)sulfonyl)-2,3-dihydro-1H-inden-1-ol(Compound 481) and(S)-2,2-difluoro-4-((1R,3S)-3-hydroxycyclohexyl)-7-((trifluoromethyl)sulfonyl)-2,3-dihydro-1H-inden-1-ol(Compound 482) and(S)-2,2-difluoro-4-((1S,3R)-3-hydroxycyclohexyl)-7-(methylsulfonyl)-2,3-dihydro-1H-inden-1-ol(Compound 483)

A solution of3-[(1S)-2,2-difluoro-1-hydroxy-7-(trifluoromethylsulfonyl)indan-4-yl]cyclohex-2-en-1-one(60.0 mg, 0.15 mmol) in methanol (3.0 mL) at 0° C. was treated withsodium borohydride (11.5 mg, 0.30 mmol) and stirred at 0° C. for 1 h.The reaction mixture was quenched by the addition of 0.5 mL of saturatedNH₄Cl. Volatiles were removed by concentration under reduced pressure.The reaction mixture was poured into 30 mL of water and extracted with3×10 mL EtOAc. The combined organics were rinsed with 10 mL of brine,dried with MgSO₄, filtered, and concentrated to dryness. Initialpurification was achieved by chromatography on silica using 10-40%EtOAc/CH₂Cl₂ to isolate 2 components. A second purification wasnecessary on the first eluting component by chromatography on silicausing 20-45% EtOAc/hexanes. Finally, each product was purifiedindividually, as described in the characterization section. Data for(S)-2,2-difluoro-4-((1R,3R)-3-hydroxycyclohexyl)-7-((trifluoromethyl)sulfonyl)-2,3-dihydro-1H-inden-1-ol(Compound 480): Final purification was achieved by chromatography onsilica using 30-70% EtOAc/hexanes to afford the desired product as awhite solid (1.5 mg, 2%). Retention time HPLC (14 min)=4.69 min; LCMSESI (−) (M+HCO₂ ⁻) m/z 445; ¹H NMR (400 MHz, CDCl₃): δ 7.92 (d, 1H),7.52 (d, 1H), 5.40 (dd, 1H), 4.33-4.28 (m, 1H), 3.59 (ddd, 1H), 3.49 (t,1H), 3.22-3.13 (m, 2H), 1.97-1.82 (m, 4H), 1.75-1.58 (m, 3H), 1.46 (dd,1H), 1.42-1.37 (m, 1H). Data for(1S)-2,2-difluoro-4-(3-hydroxycyclohex-1-en-1-yl)-7-((trifluoromethyl)sulfonyl)-2,3-dihydro-1H-inden-1-ol(Compound 481): Final purification was achieved by chromatography onsilica using 30-70% EtOAc/hexanes to afford the desired product as aclear solid (3.5 mg, 6%). Retention time HPLC (14 min)=4.70 min; LCMSESI (+) (M-OH) m/z 381; ¹H NMR (400 MHz, CDCl₃): δ 7.93 (d, 1H), 7.50(d, 1H), 5.91-5.88 (m, 1H), 5.39 (dd, 1H), 4.49-4.43 (m, 1H), 3.64 (ddd,1H), 3.39 (t, 1H), 3.18 (dd, 1H), 2.49-2.39 (m, 1H), 2.22-2.11 (m, 1H),2.09-1.92 (m, 2H), 1.81-1.59 (m, 3H). Data for(S)-2,2-difluoro-4-((1R,3S)-3-hydroxycyclohexyl)-7-((trifluoromethyl)sulfonyl)-2,3-dihydro-1H-inden-1-ol(Compound 482): Final purification was achieved by chromatography onsilica using 30-70% EtOAc/hexanes to afford the desired product as aclear solid (6.7 mg, 11%). Retention time HPLC (14 min)=4.27 min; LCMSESI (−) (M+HCO₂ ⁻) m/z 445; ¹H NMR (400 MHz, CDCl₃): δ 7.94 (d, 1H),7.55 (d, 1H), 5.41 (dd, 1H), 3.82-3.72 (m, 1H), 3.56 (ddd, 1H), 3.42 (t,1H), 3.20 (dd, 1H), 2.67 (tt, 1H), 2.17-2.08 (m, 2H), 2.01-1.94 (m, 1H),1.82-1.75 (m, 1H), 1.60-1.42 (m, 3H), 1.40-1.27 (m, 2H). Data for(S)-2,2-difluoro-4-((1S,3R)-3-hydroxycyclohexyl)-7-(methylsulfonyl)-2,3-dihydro-1H-inden-1-ol(Compound 483): Final purification was achieved by chromatography onsilica using 20-60% EtOAc/hexanes to afford the desired product as awhite solid (8.6 mg, 14%). Retention time HPLC (14 min)=4.77 min; LCMSESI (−) (M+HCO₂ ⁻) m/z 445; ¹H NMR (400 MHz, CDCl₃): δ 7.94 (d, 1H),7.55 (d, 1H), 5.41 (dd, 1H), 3.82-3.72 (m, 1H), 3.58 (ddd, 1H), 3.40 (t,1H), 3.18 (d, 1H), 2.67 (tt, 1H), 2.16-2.06 (m, 2H), 2.02-1.95 (m, 1H),1.86-1.78 (m, 1H), 1.58-1.28 (m, 5H).

Example 15:(S)-3-((2,2-difluoro-1-hydroxy-7-((trifluoromethyl)sulfonyl)-2,3-dihydro-1H-inden-4-yl)amino)-5-fluorobenzonitrile(Compound 489)

Step A: Preparation of 4-bromophenyl 3-chloropropanoate

A solution of 4-bromophenol (45.0 g, 260 mmol) in dichloromethane (1.0L) was cooled to 0° C., treated with triethylamine (44.7 g, 442 mmol). Asolution of 3-chloropropionyl chloride (36.3 g, 286 mmol) dissolved indichloromethane (100 mL) was added dropwise to the reaction vessel. Thereaction mixture was allowed to warm to ambient temperature and stirredovernight. Saturated NaCl was added to the reaction mixture, (300 mL).After stirring for 1 hour, the organic layer was separated. The aqueouslayer was extracted with dichloromethane. The combined organics werewashed with saturated NaCl, dried over Na₂SO₄, and concentrated invacuo. The crude product was used without further purification.

Step B: Preparation of 4-bromo-7-hydroxy-2,3-dihydro-1H-inden-1-one

A flask containing crude (4-bromophenyl) 3-chloropropanoate (68.0 g, 258mmol) was cooled to 0° C., then treated in several portions withaluminum trichloride (275 g, 2060 mmol). The reaction mixture was thenheated at 155° C. under N₂ for 3 hours. Stirring became difficult as thereaction proceeded. HCl (g) which was generated from the reaction wastrapped by a beaker containing 1 N NaOH. After cooling to ambienttemperature, the reaction mixture was further cooled in an ice bath.Water was added very carefully (dropwise initially and then added insmall volumes) to the reaction to quench excess AlCl₃. The mixture wasthen extracted with twice with ethyl acetate. The combined organiclayers were washed with water and brine, dried and concentrated. Thecrude product was used without additional purification.

Step C: Preparation of O-(7-bromo-3-oxo-2,3-dihydro-1H-inden-4-yl)dimethylcarbamothioate

a mixture of 4-bromo-7-hydroxy-2,3-dihydro-1H-inden-1-one (900 mg, 4.0mmol) dissolved in DMF (15 mL) was treated with DABCO 33LV (1.3 mL, 12mmol) and N,N-dimethylcarbamothioyl chloride (1.5 g, 12 mmol) wasstirred overnight at ambient temperature. The reaction was treated withwater and ethyl acetate and separated. The aqueous layer was extractedwith ethyl acetate then the combined organic layers were washed withwater and saturated NaCl. After drying, the organic layer wasconcentrated in vacuo and purified by chromatography on SiO2 elutingwith a gradient of ethyl acetate/hexanes, (670 mg, 54%). ¹H NMR (400MHz, CDCl₃): δ 7.78-7.76 (d, 1H), 6.97-6.95 (d, 1H), 3.44 (s, 3H), 3.41(s, 3H), 3.08 (m, 2H), 2.76-2.69 (m, 2H).

Step D: Preparation of S-(7-bromo-3-oxo-2,3-dihydro-1H-inden-4-yl)dimethylcarbamothioate

A mixture of O-(7-bromo-3-oxo-2,3-dihydro-1H-inden-4-yl)dimethylcarbamothioate (670 mg, 2.1 mmol) and diphenyl ether (15 mL) washeated at 220° C. under N₂ for 30 minutes. After cooling to ambienttemperature, the mixture was diluted with hexanes and the mixture wasapplied to a pad of SiO₂ and eluted with hexanes. After removal of thediphenyl ether, the desired product was eluted with ethyl acetate. Afterconcentration in vacuo, the crude product was used without furtherpurification.

Step E: Preparation of 4-bromo-7-mercapto-2,3-dihydro-1H-inden-1-one

A solution of S-(7-bromo-3-oxo-2,3-dihydro-1H-inden-4-yl)dimethylcarbamothioate (670 mg, 2.1 mmol) dissolved in ethanol (25 mL)was treated with 3N sodium hydroxide) 10.7 mL, 32.1 mmol). The mixturewas heated to reflux for 1 hour then cooled to 0° C. Aqueous HCl (3M)was added dropwise to neutralize the reaction. Ethanol was removed byconcentration in vacuo followed by addition of aqueous HCl (1M) toadjust to pH 3-4. The aqueous was extracted twice with ethyl acetate andthe combined organic layers were washed with saturated NaCl, dried andconcentrated in vacuo. The crude product was used without furtherpurification.

Step F: Preparation of4-bromo-7-((trifluoromethyl)thio)-2,3-dihydro-1H-inden-1-one

Methyl viologen dichloride hydrate (0.11 g, 0.41 mmol),4-bromo-7-mercapto-2,3-dihydro-1H-inden-1-one (2.0 g, 8.2 mmol) andtriethylamine (1.25 g, 12.3 mmol) were dissolved in DMF (50 mL) andcooled to −50° C. The flask was placed under gentle vacuum thentrifluoromethyl iodide (3.2 g, 16 mmol) gas was introduced using aballoon. This reaction was warmed to ambient temperature and stirred atovernight. The reaction mixture was diluted with ethyl acetate andwater, filtered through a celite pad, and the layers were partitioned.The organic layer was washed with water, brine, dried over Na₂SO₄,filtered, and evaporated. The crude oil was then purified by flashcolumn chromatography on SiO₂ eluting with petroleum ether/ethylacetate, (0.96 g, 51.7%). ¹H NMR (400 MHz, CDCl₃): δ 7.72 (d, 1H), 7.41(d, 1H), 3.10-3.07 (m, 2H), 2.79-2.77 (m, 2H).

Step G: Preparation of4-bromo-7-((trifluoromethyl)sulfonyl)-2,3-dihydro-1H-inden-1-one

Ruthenium(III) chloride (19 mg, 0.09 mmol) was added to a mixture of4-bromo-7-((trifluoromethyl)thio)-2,3-dihydro-1H-inden-1-one (0.96 g,3.1 mmol) and sodium periodate (1.98 g, 9.26 mmol) in a mixture ofcarbon tetrachloride (20 mL), acetonitrile (20 mL), and water (40 mL).The mixture was stirred at ambient temperature for 3 hours. The reactionmixture was partitioned between dichloromethane and water. The organiclayer was washed with brine, dried over MgSO₄, filtered, andconcentrated in vacuo. The crude product was purified by flash columnchromatography on SiO₂ eluting with petroleum ether/ethyl acetate, (1.7g, 79%). ¹H NMR (400 MHz, CDCl₃): δ 8.05-8.02 (m, 2H), 3.21-3.18 (m,2H), 2.89-2.86 (m, 2H).

Step H: Preparation of4-bromo-7-((trifluoromethyl)sulfonyl)-2,3-dihydrospiro[indene-1,2′-[1,3]dioxolane]

Trimethylsilyl trifluoromethanesulfonate (177 mg, 0.80 mmol) was addeddropwise to a pre-cooled (−78° C.) solution of4-bromo-7-((trifluoromethyl)sulfonyl)-2,3-dihydro-1H-inden-1-one andtrimethyl(2-trimethylsilyloxyethoxy)silane (410 mg, 2.0 mmol) dissolvedin dichloromethane (50 mL). The reaction mixture was warmed to ambienttemperature and stirred for 2 hours. The reaction was quenched byaddition of triethylamine then concentrated in vacuo. The residue wasredissolved in ethyl acetate and washed twice with water, and saturatedNaCl. The organic layer was separated, dried over Na₂SO₄ andconcentrated in vacuo. The crude product was purified by chromatographyon SiO₂ eluting with ethyl acetate/isohexane, (600 mg, 77%).

Step I: Preparation of4-bromo-7-((trifluoromethyl)sulfonyl)-2,3-dihydro-1H-inden-1-one

4-Bromo-7-((trifluoromethyl)sulfonyl)-2,3-dihydrospiro[indene-1,2′-[1,3]dioxolane](3.5 g, 9.1 mmol) was dissolved in THF (72 mL) and treated with 10%aqueous HCl (27 mL, 27 mmol). The mixture was stirred for severalminutes then warmed to 60° C. for 2 hours. The mixture was cooled,diluted with diethyl ether and separated. The aqueous was washed withdiethyl ether and the combined organics were washed with water,saturated NaHCO₃, saturated NaCl, dried over Na₂SO₄ and concentrated invacuo to a yellowish solid, (3.09 g, quant.). ¹H NMR (400 MHz, CDCl₃): δ8.05-8.02 (m, 2H), 3.21-3.18 (m, 2H), 2.89-2.86 (m, 2H).

Step J: Preparation of(E.Z)-3-((4-bromo-7-((trifluoromethyl)sulfonyl)-2,3-dihydro-1H-inden-1-ylidene)amino)propan-1-ol

4-Bromo-7-(trifluoromethylsulfonyl)indan-1-one (3.09 g, 9.02 mmol] wasslurried in toluene (35 mL) and cyclohexane (35 mL) then treated with3-methoxypropylamine (2.15 mL, 27.1 mmol) and pivalic acid (46 mg, 0.45mmol). The mixture was refluxed through a Dean-Stark trap (sidearmpre-filled with cyclohexane) for 8 hours. The reaction mixture wascooled and concentrated in vacuo. The crude material was taken directlyinto the fluorination.

Step K: Preparation of4-bromo-2,2-difluoro-7-((trifluoromethyl)sulfonyl)-2,3-dihydro-1H-inden-1-one

Crude(E.Z)-3-((4-bromo-7-((trifluoromethyl)sulfonyl)-2,3-dihydro-1H-inden-1-ylidene)amino)propan-1-ol(3.75 g, 9.1 mmol) was dissolved in dry acetonitrile (23 mL) and addeddropwise to a warm (60° C.), suspension of Selectfluor (9.6 g, 27.2mmol) and sodium sulfate (12.9 g, 90.5 mmol) slurried in acetonitrile(10 mL). After the addition, the mixture was heated to 60° C. for 10minutes then cooled to ambient temperature and treated with 10% HCl (15mL) and stirred for 20 minutes. The mixture was adjusted to pH 8 withsolid NaHCO₃ then diluted with ethyl acetate and separated. The aqueouswas washed with ethyl acetate and the combined organics were washed withsaturated NaHCO₃, saturated NaCl, dried over Na₂SO₄ filtered, andconcentrated in vacuo to dark oil. The crude material waschromatographed on SiO₂ eluting with a gradient of ethylacetate/hexanes. The desired product was concentrated to a light yellowsolid, (2.27 g, 66%). ¹H NMR (400 MHz, CDCl₃): δ 8.22-8.14 (m, 2H),3.60-3.55 (t, 2H).

Step L: Preparation of(S)-4-bromo-2,2-difluoro-7-((trifluoromethyl)sulfonyl)-2,3-dihydro-1H-inden-1-ol

4-Bromo-2,2-difluoro-7-((trifluoromethyl)sulfonyl)-2,3-dihydro-1H-inden-1-one(1.65 g, 4.35 mmol) was dissolved in isopropanol (21 mL) and treatedwith triethylamine (1.2 mL, 8.7 mmol), formic acid (0.49 mL, 13.1 mmol)and RuCl(p-cymene)[(R,R)-Ts-DPEN] (27.7 mg, 0.040 mmol). The reactionmixture was stirred at ambient temperature for 4 hours. The solvent wasremoved in vacuo then the crude material was chromatographed on SiO₂eluting with a gradient of ethyl acetate/hexanes. The product wasisolated as a more pure fraction (1.83 g) and a slightly less purefraction. Both of these fractions were successfully utilized in thecoupling reaction. ¹H NMR (400 MHz, CDCl₃): δ 7.88-7.80 (m, 2H),5.50-5.45 (m, 1H), 3.66-3.58 (m, 1H), 3.20 (m, 1H).

Step M: Preparationof(S)-3-((2,2-difluoro-1-hydroxy-7-((trifluoromethyl)sulfonyl)-2,3-dihydro-1H-inden-4-yl)amino)-5-fluorobenzonitrile

(S)-4-Bromo-2,2-difluoro-7-((trifluoromethyl)sulfonyl)-2,3-dihydro-1H-inden-1-ol(98 mg, 0.26 mmol) was dissolved in 1,4-dioxane (0.80 mL) and treatedwith benzonitrile, 3-amino-5-fluoro-(42 mg, 0.31 mmol), palladium (II)acetate (2.9 mg, 0.010 mmol), and Xantphos (14.9 mg, 0.030 mmol). Themixture was heated to 120° C. for 1.5 hours in the microwave reactor.The reaction mixture was cooled, diluted with ethyl acetate and waterthen separated. The aqueous was washed with ethyl acetate and thecombined organics were washed with saturated NaHCO₃, saturated NaCl,dried over Na₂SO₄ and concentrated in vacuo. The crude dark oil waschromatographed on SiO₂ eluting with a gradient of ethylacetate/hexanes. The desired material was recovered in a slightly impureform. This material was re-chromatographed on reversed-phase SiO₂eluting with a gradient of MeCN/water. A single fraction was collectedand to light tan solid, (35 mg, 31%). LCMS ESI (−) m/z (M−H) 435; ¹H NMR(400 MHz, CDCl₃): δ 7.87 (d, 1H), 7.31-7.29 (m, 2H), 7.21-7.19 (m, 2H),6.18 (m, 1H), 5.42-5.38 (m, 1H), 3.52-3.41 (m, 1H), 3.32-3.24 (m, 1H).

Example 16: Synthesis of(S)-5-((2,2-difluoro-1-hydroxy-7-((trifluoromethyl)sulfonyl)-2,3-dihydro-1H-inden-4-yl)amino)nicotinonitrile(Compound 488)

(S)-4-Bromo-2,2-difluoro-7-((trifluoromethyl)sulfonyl)-2,3-dihydro-1H-inden-1-ol(23 mg, 0.060 mmol) was dissolved in 1,4-dioxane (0.20 mL) and treatedwith 3-pyridinecarbonitrile, 5-amino- (8.6 mg, 0.070 mmol), cesiumcarbonate (27.5 mg, 0.080 mmol), palladium (II) acetate (0.68 mg, 0.003mmol), and Xantphos (3.5 mg, 0.010 mmol). After cooling, the mixture wasdiluted with water and ethyl acetate then separated. This mixture didn'tseparate well and there was insoluble yellow solid present, which wasremoved by filtration. The aqueous was washed with ethyl acetate and thecombined organics were washed with saturated NaHCO₃, saturated NaCl,dried over Na2SO4, and concentrated in vacuo to a dark residue. Thecrude material was chromatographed on SiO₂ eluting with a gradient ofethyl acetate/hexanes. The product was recovered as light tan solid, (12mg, 47%). LCMS ESI (+) m/z (M+H) 420; ¹H NMR (400 MHz, CDCl₃ plusCD₃OD): δ 8.66 (s, 1H), 8.60 (s, 1H), 7.79-7.75 (m, 2H), 7.22-7.20 (m,1H), 5.30 (d, 1H), 3.92-3.90 (m, 1H), 3.46-3.32 (m, 1H), 3.31-3.21 (m,1H).

Example 17: Synthesis of(S)-2,2-difluoro-4-morpholino-7-((trifluoromethyl)sulfonyl)-2,3-dihydro-1H-inden-1-ol(Compound 490)

Step A: Preparation of(S)-((4-bromo-2,2-difluoro-7-((trifluoromethyl)sulfonyl)-2,3-dihydro-1H-inden-1-yl)oxy)(tert-butyl)dimethylsilane

(S)-4-bromo-2,2-difluoro-7-((trifluoromethyl)sulfonyl)-2,3-dihydro-1H-inden-1-ol(0.30 g, 0.78 mmol) was dissolved in methylene chloride (3.3 mL) andtreated with 2,6-lutidine (0.40 mL, 3.1 mmol), cooled to 0° C., followedby treatment with t-butyldimethylsilyl triflate (0.45 mL, 1.9 mmol). Themixture was warmed to ambient temperature and stirred for two hours. Thereaction mixture was re-cooled to 0° C. and cold 10% KHSO₄ was addedalong with additional methylene chloride then the layers were separated.The organic layer was washed with 10% KHSO₄, water, then with one-halfsaturated NaHCO₃. The organic layer was dried over Na₂SO₄ andconcentrated in vacuo to a light yellow oil. The crude product waschromatographed on SiO₂ eluting with a gradient of methylenechloride/hexanes. The product was concentrated to colorless oil, (466mg, 92%).

Step B: Preparation of(S)-4-(1-((tert-butyldimethylsilyl)oxy)-2,2-difluoro-7-((trifluoromethyl)sulfonyl)-2,3-dihydro-1H-inden-4-yl)morpholine

(S)-((4-bromo-2,2-difluoro-7-((trifluoromethyl)sulfonyl)-2,3-dihydro-1H-inden-1-yl)oxy)(tert-butyl)dimethylsilane(0.030 g, 0.060 mmol) was dissolved in DMF (0.25 mL) and treated withsodium acetate (14 mg, 0.17 mmol) followed by morpholine (0.020 mL, 0.17mmol). The mixture was heated at 120° C. for 1 hour in the microwavereactor. After cooling, the mixture was diluted with ethyl acetate thenwashed 7 times with water, saturated NaCl, dried over Na₂SO₄ andconcentrated in vacuo. The residue was chromatographed on SiO₂ elutingwith a gradient of ethyl acetate/hexanes. The product was recovered as acolorless film, (17 mg, 58%). LCMS ESI (+) m/z (M+H) 502.

Step C: Preparation of(S)-2,2-difluoro-4-morpholino-7-((trifluoromethyl)sulfonyl)-2,3-dihydro-1H-inden-1-ol

(S)-4-(1-((tert-butyldimethylsilyl)oxy)-2,2-difluoro-7-((trifluoromethyl)sulfonyl)-2,3-dihydro-1H-inden-4-yl)morpholine(0.02 g, 0.04 mmol) was dissolved in THF (0.25 mL) and treated withglacial acetic acid (2 μL, 0.04 mmol) followed by a solution of 1 Mtetrabutylammonium fluoride in THF (0.04 mL, 0.04 mmol). The mixture washeated to 60° C. for 45 minutes. The darkened reaction mixture wascooled and added into saturated aqueous NaHCO₃. The mixture was vortexedvigorously then diluted with ethyl acetate and separated. The organiclayer was washed with saturated NaHCO₃, saturated NaCl, dried overNa₂SO₄ and concentrated in vacuo. The crude material was chromatographedon SiO₂ eluting with a gradient of ethyl acetate/hexanes. Compound 490was recovered as light orange oil, (10.8 mg, 78%). LCMS ESI (+) m/z(M+H) 388; ¹H NMR (400 MHz, CDCl₃): δ 7.87 (d, 1H), 7.01 (d, 1H),5.35-5.31 (m, 1H), 3.93-3.81 (m, 4H), 3.56-3.45 (m, 1H), 3.35-3.23 (m,3H), 3.16-3.09 (m, 3H).

Example 18: Synthesis of(1S,3R)-4-((3-chloro-5-fluorophenyl)thio)-2,2,3-trifluoro-7-(methylsulfonyl)-2,3-dihydro-1H-inden-1-ol(Compound 491)

Step A: Preparation of 4,7-difluoro-1H-indene-1,3(2H)-dione

(0.52 g, 2.8 mmol) was slurried acetic anhydride (2.5 mL, 27 mmol) andtreated with tert-butyl 3-oxobutanoate (0.52 mL, 3.1 mmol) andtriethylamine (1.4 mL, 10 mmol). The mixture was stirred at ambienttemperature for 60 hours. The reaction was cooled to 0° C. and treatedwith 10% aqueous hydrochloric acid (8.6 mL, 25 mmol) by dropwiseaddition. After the addition, the mixture was warmed to ambienttemperature then heated to 75° C. for 10 minutes. After cooling, themixture was diluted with water (20 mL) and extracted three times withmethylene chloride (20 mL portions). The combined organics were driedover Na₂SO₄ and concentrated in vacuo to crude orange solid. Thismaterial was carried forward without purification.

Step B: Preparation of 2,2,4,7-tetrafluoro-1H-indene-1,3(2H)-dione

4,7-difluoro-1H-indene-1,3(2H)-dione (0.51 g, 2.8 mmol) was dissolved inacetonitrile (27 mL), placed in an ambient temperature water bath thentreated with solid sodium carbonate (950 mg, 9.0 mmol) followed bySelectfluor® (2.18 g, 6.2 mmol). The mixture was stirred at ambienttemperature for 1 hour. The mixture was filtered to removed undissolvedsolids, the solids were washed with ethyl acetate and the filtrate wasconcentrated in vacuo. The residue was redissolved in water (ca. 20 mL)and extracted four times with ethyl acetate (20 mL each). The combinedorganics were washed with saturated NaCl, dried over Na₂SO₄ andconcentrated in vacuo to orange solid. The crude solid waschromatographed on SiO₂ eluting with an aggressive gradient of ethylacetate/hexanes. The desired material concentrated to orange solid, (493mg, 81%). ¹H NMR (400 MHz, CDCl₃): δ 7.70-7.65 (2H).

Step C: Preparation of(S)-2,2,4,7-tetrafluoro-3-hydroxy-2,3-dihydro-1H-inden-1-one

2,2,4,7-tetrafluoro-1H-indene-1,3 (2H)-dione (5.81 g, 26.6 mmol) wassuspended in methylene chloride (260 mL), cooled to 0° C., and treatedwith formic acid (1.01 mL, 26.6 mmol), triethylamine (2.60 mL, 18.6mmol), then the reaction mixture was sparged with argon for 5 minutes.RuCl(p-cymene)[(S,S)-Ts-DPEN] (339 mg, 0.530 mmol) was added and thereaction was transferred to the refrigerator and allowed to stand at 4°C. for 20 hours. The cold reaction mixture was poured into cold 1Naqueous HCl (70 mL) and separated. The aqueous was washed twice withethyl acetate then the combined organic layers were dried over Na₂SO₄and concentrated in vacuo to a brown semi-solid. The crude material waschromatographed on SiO₂ eluting with a gradient of ethylacetate/hexanes. The product was recovered as yellow solid, (3.48 g,59%). ¹H NMR (400 MHz, CDCl₃): δ 7.86-7.80 (m, 1H), 7.60-7.54 (m, 1H),5.79-5.74 (m, 1H), 3.23-3.18 (m, 1H).

Step D: Preparation of(S)-2,2,4-trifluoro-3-hydroxy-7-(methylthio)-2,3-dihydro-1H-inden-1-one

(S)-2,2,4,7-tetrafluoro-3-hydroxy-2,3-dihydro-1H-inden-1-one (0.40 g,1.8 mmol) was dissolved in dry acetonitrile (18 mL), cooled to 0° C.,and sparged with argon for 5 minutes. The solution was treated in asingle portion with sodium thiomethoxide (144 mg, 2.06 mmol) and after 5minutes, the ice bath was removed and the reaction was stirred atambient temperature for 3 hours. The reaction mixture was concentratedin vacuo and the residue was redissolved in water and ethyl acetate.After separation, the aqueous was washed twice with ethyl acetate andthe combined organics were washed with saturated NaCl, dried over Na₂SO₄and concentrated in vacuo. The orange residue was chromatographed onSiO₂ eluting with a gradient of ethyl acetate/hexanes. The desiredmaterial was recovered as bright yellow solid, (314 mg, 70%). LCMS ESI(+) m/z (M+H) 249.

Step E: Preparation of(S)-2,2,4-trifluoro-3-hydroxy-7-(methylsulfonyl)-2,3-dihydro-1H-inden-1-one

(S)-2,2,4-trifluoro-3-hydroxy-7-(methylthio)-2,3-dihydro-1H-inden-1-one(0.40 g, 1.6 mmol) was dissolved in MeOH (10 mL) and the reaction wastreated dropwise with a solution of Oxone® (2.2 g, 3.6 mmol) dissolvedin water (10 mL). The mixture was stirred at ambient temperature for 14hours. The reaction mixture was filtered, the solids were washed withethyl acetate and the filtrate was concentrated in vacuo to removevolatile solvents. The aqueous filtrate was extracted three times withethyl acetate then the combined organics were washed with saturatedNaCl, dried over Na₂SO₄ and concentrated in vacuo to yellow solid, (467mg, quant.). LCMS ESI (+) m/z (M+H) 281.

Step F: Preparation of(R)-2,2,3,4-tetrafluoro-7-(methylsulfonyl)-2,3-dihydro-1H-inden-1-one

(S)-2,2,4-trifluoro-3-hydroxy-7-(methylsulfonyl)-2,3-dihydro-1H-inden-1-one(0.45 g, 1.6 mmol) was dissolved in dichloromethane (16 mL), cooled to0° C., and treated dropwise with diethylaminosulfur trifluoride (DAST)(0.32 mL, 2.4 mmol) and stirred at 0° C. for 14 hours. The reaction wastreated with additional diethylaminosulfur trifluoride (0.32 mL, 2.4mmol) and stirring continued for 6 hours at 0° C. The cold reaction wastreated with saturated NaHCO₃ (10 mL) and stirred vigorously for 20minutes. The mixture was diluted with additional methylene chloride andthe layers were separated. The aqueous was re-extracted with methylenechloride and the combined organic layers were dried over Na₂SO₄ andconcentrated in vacuo to a yellow solid. The crude material waschromatographed on SiO₂ eluting with a gradient of ethylacetate/hexanes. The desired material was recovered as pale yellowsolid, (258 mg, 53%). LCMS ESI (+) m/z (M+H) 283.

Step G: Preparation of(1S,3R)-2,2,3,4-tetrafluoro-7-(methylsulfonyl)-2,3-dihydro-1H-inden-1-ol

(R)-2,2,3,4-tetrafluoro-7-(methylsulfonyl)-2,3-dihydro-1H-inden-1-one:(0.098 g, 0.35 mmol) was suspended in methylene chloride (3.3 mL),cooled to 0° C., and treated with triethylamine (97 μL, 0.69 mmol),formic acid (39 μL, 1.0 mmol) and RuCl(p-cymene)[(R,R)-Ts-DPEN] (2.2 mg,0.003 mmol). The solution was allowed to stand at 4° C. in therefrigerator for 60 hours. The reaction mixture was concentrated in astream of nitrogen gas then chromatographed on SiO₂ eluting with agradient of ethyl acetate/hexanes. The desired fractions wereconcentrated to colorless film, (53 mg, 53%). LCMS ESI (+) m/z (M+H)285.

Step H: Preparation of(1S,3R)-4-((3-chloro-5-fluorophenyl)thio)-2,2,3-trifluoro-7-(methylsulfonyl)-2,3-dihydro-1H-inden-1-ol

(1S,3R)-2,2,3,4-tetrafluoro-7-(methylsulfonyl)-2,3-dihydro-1H-inden-1-ol(0.005 g, 0.02 mmol) was treated with cesium bicarbonate (17 mg, 0.090mmol) and suspended in DMF (0.1 mL) then stirred at ambient temperaturefor 1 hour. 3-Chloro-5-fluorothiophenol (14 mg, 0.090 mmol) was addedand the mixture was stirred at ambient temperature for 18 hours. Thereaction was concentrated in a stream of nitrogen gas to remove DMF. Theresidue was chromatographed on SiO₂ eluting with a stepped gradient ofethyl acetate/hexanes. Compound 491 was concentrated to light pink oil,(7 mg, 93%). LCMS ESI (+) m/z (M+Na) 449; ¹H NMR (400 MHz, CDCl₃): δ7.99-7.95 (m, 1H), 7.33-7.32 (m, 1H), 7.24-7.19 (m, 2H), 7.16-7.13 (m,1H), 5.75 (dd, 1H), 5.68-5.65 (m, 1H), 3.37-3.36 (m, 1H), 3.23 (s, 3H).

Example 19: Synthesis of3-(((1S,3R)-2,2,3-trifluoro-1-hydroxy-7-(methylsulfonyl)-2,3-dihydro-1H-inden-4-yl)thio)benzonitrile(Compound 492)

(1S,3R)-2,2,3,4-tetrafluoro-7-(methylsulfonyl)-2,3-dihydro-1H-inden-1-ol(0.0073 g, 0.030 mmol) was treated with cesium bicarbonate (25 mg, 0.13mmol) and suspended in DMF (0.1 mL) then 3-mercapto-benzonitrile (17 mg,0.13 mmol) was added and the mixture was stirred at ambient temperaturefor 60 hours. The reaction was concentrated in a stream of nitrogen gasto remove DMF. The residue was chromatographed on SiO₂ eluting with astepped gradient of ethyl acetate/hexanes. The product was concentratedto light oil, (7 mg, 91%). LCMS ESI (+) m/z (M+Na) 422; ¹H NMR (400 MHz,CDCl₃): δ 7.97-7.87 (m, 1H), 7.81 (m, 1H), 7.77-7.74 (m, 2H), 7.61-7.57(m, 1H), 7.16-7.14 (m, 1H), 5.77 (dd, 1H), 5.69-5.65 (m, 1H), 3.40-3.39(m, 1H), 3.23 (s, 3H).

Example 20: Synthesis of(S)-4-(3-chlorophenyl)-2,2-difluoro-7-((trifluoromethyl)sulfonyl)-2,3-dihydro-1H-inden-1-ol(Compound 485)

Step A: Preparation of4-((4-methoxybenzyl)oxy)-7-((trifluoromethyl)sulfonyl)-2,3-dihydrospiro[indene-1,2′-[1,3]dioxolane]

A solution of4′-fluoro-7′-(trifluoromethylsulfonyl)spiro[1,3-dioxolane-2,1′-indane](1.0 g, 3.07 mmol) and 4-methoxybenzyl alcohol (760 μL, 6.13 mmol) inacetonitrile (9.3 mL) at 25° C. was treated with potassium hydroxide(516 mg, 9.2 mmol) and stirred at 25° C. for 1.5 h. The reaction mixturewas poured into 150 mL of water and extracted with 3×40 mL EtOAc. Thecombined organics were rinsed with 20 mL of brine, dried with MgSO₄,filtered, and concentrated to dryness. Purification was achieved bychromatography on silica using 10-25% EtOAc/hexane to afford4-((4-methoxybenzyl)oxy)-7-((trifluoromethyl)sulfonyl)-2,3-dihydrospiro[indene-1,2′-[1,3]dioxolane]as a solid (1.08 g, 79%). LCMS ESI (+) [M+H]⁺ m/z 445.

Step B: Preparation of4-((4-methoxybenzyl)oxy)-7-((trifluoromethyl)sulfonyl)-2,3-dihydro-1H-inden-1-one

In a glass pressure vessel, a solution of4′-[(4-methoxyphenyl)methoxy]-7′-(trifluoromethylsulfonyl)spiro[1,3-dioxolane-2,1′-indane](1.08 g, 2.43 mmol) in a mixture acetone (20 mL) and water (20 mL) wastreated with pyridinium p-toluenesulfonate (122 mg, 0.49 mmol), sealedand stirred at 80° C. overnight. Volatiles were removed by concentrationunder reduced pressure. The residue was poured into 40 mL of saturatedaqueous NaHCO₃ and extracted with 3×50 mL EtOAc. The combined organicswere rinsed with 30 mL of brine, dried with MgSO₄, filtered, andconcentrated to dryness. The product was used without furtherpurification. LCMS ESI (+) [M+H]⁺ m/z 401.

Step C: Preparation of2,2-difluoro-4-((4-methoxybenzyl)oxy)-7-((trifluoromethyl)sulfonyl)-2,3-dihydro-1H-inden-1-one

2,2-Dimethylpropanoic acid (50 mg, 0.46 mmol) was added to a flaskcontaining a suspension of4-[(4-methoxyphenyl)methoxy]-7-(trifluoromethylsulfonyl)indan-1-one (922mg, 2.3 mmol) and 3-methoxypropan-1-amine (0.35 mL, 3.45 mmol) in amixture of toluene (14 mL) and cyclohexane (14 mL). This was refluxedwith a Dean-Stark trap attached at 104° C. After 2.5 h, the reactionmixture was cooled and volatiles removed by concentration under reducedpressure. The residue was dissolved in acetonitrile (24 mL) and treatedsequentially with sodium sulfate (850 mg, 6.0 mmol) and1-(chloromethyl)-4-fluoro-1,4-diazoniabicyclo[2.2.2]octaneditetrafluoroborate (2.13 g, 6.0 mmol). The resulting suspension stirredat 60° C. for 2 h. After cooling to room temperature, the reactionmixture was treated with concentrated hydrochloric acid (600 μL, 7.2mmol) and water (10 mL). The resulting mixture stirred for 20 min.Volatiles were removed by concentration under reduced pressure. Thereaction mixture was poured into 20 mL of water and extracted with 3×30mL EtOAc. The combined organics were rinsed with 20 mL of brine, driedwith MgSO₄, filtered, and concentrated to dryness. Purification wasachieved by chromatography on silica using 5-40% EtOAc/hexane to afford2,2-difluoro-4-((4-methoxybenzyl)oxy)-7-((trifluoromethyl)sulfonyl)-2,3-dihydro-1H-inden-1-oneas a solid (520 mg, 50%). LCMS ESI (+) [M+H]⁺ m/z 437.

Step D: Preparation of(S)-2,2-difluoro-4-((4-methoxybenzyl)oxy)-7-((trifluoromethyl)sulfonyl)-2,3-dihydro-1H-inden-1-ol

a solution of2,2-difluoro-4-[(4-methoxyphenyl)methoxy]-7-(trifluoromethylsulfonyl)indan-1-one(520 mg, 1.19 mmol) in dichloromethane (11.9 mL) was cooled to 0° C. andsparged with nitrogen for 5 minutes. During this time, formic acid (130μL, 3.58 mmol) and triethylamine (330 μL, 2.38 mmol) were sequentiallyadded. Once the sparging was complete,RuCl(p-cymene)[(R,R)-Ts-DPEN](22.8 mg, 0.036 mmol) was added under acontinuous stream of nitrogen. The reaction vessel was sealed and putinto the refrigerator to react overnight. Once complete, the reactionmixture was poured into 30 mL of saturated aqueous NaHCO₃ and extractedwith 3×30 mL CH₂Cl₂. The combined organics were rinsed with 20 mL ofbrine, dried with MgSO₄, filtered, and concentrated to dryness.Purification was achieved by chromatography on silica using 10-40%EtOAc/hexane to afford(S)-2,2-difluoro-4-((4-methoxybenzyl)oxy)-7-((trifluoromethyl)sulfonyl)-2,3-dihydro-1H-inden-1-olas a solid (370 mg, 71%). LCMS ESI (+) [M+NH₄]⁻ m/z 456.

Step E: Preparation of(S)-tert-butyl((2,2-difluoro-4-((4-methoxybenzyl)oxy)-7-((trifluoromethyl)sulfonyl)-2,3-dihydro-1H-inden-1-yl)oxy)dimethylsilane

A solution of(1S)-2,2-difluoro-4-[(4-methoxyphenyl)methoxy]-7-(trifluoromethylsulfonyl)indan-1-ol(370 mg, 0.84 mmol) and 2,6-lutidine (780 μL, 7.76 mmol) indichloromethane (8.4 mL) at −78° C. was treated withtert-butyldimethylsilyl trifluoromethanesulfonate (980 μL, 4.22 mmol)and allowed to warm to room temperature over 2 h. The reaction mixturewas poured into 30 mL of saturated aqueous NaHCO₃ and extracted with3×20 mL CH₂Cl₂. The combined organics were rinsed with 10 mL of brine,dried with MgSO₄, filtered, and concentrated to dryness. Purificationwas achieved by chromatography on silica using 5-20% EtOAc/hexane toafford(S)-tert-butyl((2,2-difluoro-4-((4-methoxybenzyl)oxy)-7-((trifluoromethyl)sulfonyl)-2,3-dihydro-1H-inden-1-yl)oxy)dimethylsilane(460 mg, quant). LCMS ESI (−) [M−H]⁻ m/z 551.

Step F: Preparation of(S)-1-((tert-butyldimethylsilyl)oxy)-2,2-difluoro-7-((trifluoromethyl)sulfonyl)-2,3-dihydro-1H-inden-4-ol

A solution of tert-butyl-[(1S)-2,2-difluoro-4-[(4-methoxyphenyl)methoxy]-7-(trifluoromethylsulfonyl)indan-1-yl]oxy-dimethylsilane(460 mg, 0.83 mmol) in dichloromethane (3.0 mL) at 25° C. was treatedwith trifluoroacetic acid (3.0 mL) and stirred at 25° C. After 1 h,volatiles were removed by concentration under reduced pressure. To theresulting residue was added 6 mL of toluene and the organic volatileswere once again removed by concentration under reduced pressure. Thisprocess was repeated twice. Purification was achieved by chromatographyon reverse phase by injection of a DMF solution of the product residue.40-100% CH₃CN/Water was used as eluent.(S)-1-((tert-butyldimethylsilyl)oxy)-2,2-difluoro-7-((trifluoromethyl)sulfonyl)-2,3-dihydro-1H-inden-4-olwas isolated a thick orange oil (236 mg, 89%). LCMS ESI (−) [M−H]⁻ m/z431.

Step G: Preparation of(S)-2,2-difluoro-1-hydroxy-7-((trifluoromethyl)sulfonyl)-2,3-dihydro-1H-inden-4-yltrifluoromethanesulfonate

A solution of(S)-1-((tert-butyldimethylsilyl)oxy)-2,2-difluoro-7-((trifluoromethyl)sulfonyl)-2,3-dihydro-1H-inden-4-ol(215 mg, 0.50 mmol) and 2,6-bis(1,1-dimethylethyl)-4-methyl-pyridine(408 mg, 1.99 mmol) in dichloromethane (10 mL) was cooled to −78° C. andtreated with trifluoromethanesulfonic anhydride (0.17 mL, 0.99 mmol).The mixture was stirred at −78° C. for 1 h. The reaction mixture waspoured into 20 mL of saturated NaHCO₃ and extracted with 3×20 mL EtOAc.The combined organics were rinsed with 10 mL of brine, dried with MgSO₄,filtered, and concentrated to dryness. Purification was achieved bychromatography on SiO₂ eluting with a gradient of ethyl acetate/hexanes.

Step H: Preparation of tributyl(3-chlorophenyl)stannane

A solution of 3-chlorophenylmagnesium bromide (0.5 M in THF) (1.40 mL,0.70 mmol) was cooled to −78° C. and treated dropwise with a solution oftributyl(chloro)stannane (230 mg, 0.70 mmol) dissolved in THF (0.5 mL).The solution was stirred for 5 minutes then allowed to warm to ambienttemperature slowly without the bath and stirred at ambient temperaturefor 45 hours. The reaction was quenched by addition of saturated NH₄Cland water. Diethyl ether was added and the mixture was separated. Theaqueous was washed twice with diethyl ether and the combined organicswere washed saturated NaCl, dried over Na₂SO₄ and concentrated in vacuo.The crude material was chromatographed on SiO₂ eluting with cyclohexane.The desired product was concentrated to colorless liquid, (234 mg, 84%).¹H NMR (400 MHz, CDCl₃): δ 7.58-7.30 (m, 4H), 1.60-1.45 (m, 6H),1.40-1.28 (m, 6H), 1.17-1.00 (m, 6H), 0.92-0.85 (m, 9H).

Step I: Preparation of(S)-tert-butyl((4-(3-chlorophenyl)-2,2-difluoro-7-((trifluoromethyl)sulfonyl)-2,3-dihydro-1H-inden-1-yl)oxy)dimethylsilane

(S)-2,2-difluoro-1-hydroxy-7-((trifluoromethyl)sulfonyl)-2,3-dihydro-1H-inden-4-yltrifluoromethanesulfonate (17 mg, 0.030 mmol) was dissolved in toluene(0.4 mL) then tributyl-(3-chlorophenyl)stannane (23 mg, 0.060 mmol),tetrakis(triphenylphosphine)palladium (1.7 mg, 0.001 mmol), and lithiumchloride (4 mg, 0.09 mmol) were added. The reaction mixture was spargedwith argon then heated to reflux for 16 hours. After cooling, themixture was concentrated in a stream of nitrogen gas. The crude materialwas chromatographed on SiO₂ eluting with a gradient of ethylacetate/hexanes. The less polar UV-active spot was recovered ascolorless oil, (9.8 mg, 63%).

Step J: Preparation of(S)-4-(3-chlorophenyl)-2,2-difluoro-7-((trifluoromethyl)sulfonyl)-2,3-dihydro-1H-inden-1-ol

(S)-tert-butyl((4-(3-chlorophenyl)-2,2-difluoro-7-((trifluoromethyl)sulfonyl)-2,3-dihydro-1H-inden-1-yl)oxy)dimethylsilane(9.8 mg, 0.020 mmol) was dissolved in THF (0.2 mL) containing glacialacetic acid (3.2 μL, 0.060 mmol) and the mixture was treated with a 1Msolution of tetrabutylammonium fluoride in THF (28 μL, 0.030 mmol). Themixture was heated to 60° C. for 2 hours, then the reaction was cooled,treated with one-half saturated NaHCO₃, diluted with ethyl acetate andseparated. The aqueous was washed twice with ethyl acetate and thecombined organics were washed with saturated NaHCO₃, saturated NaCl,dried over Na₂SO₄ and concentrated in vacuo to dark oil. The crudematerial was chromatographed on SiO₂ eluting with a stepped gradient ofethyl acetate/hexanes. The desired product was concentrated to lightyellow solid, (2.5 mg, 32%). LCMS ESI (−) m/z (M−H) 411/413; ¹H NMR (400MHz, CDCl₃): δ 8.06-8.04 (d, 1H), 7.66-7.64 (d, 1H), 7.48-7.41 (m, 3H),7.29-7.26 (m, 1H), 5.47-5.43 (m, 1H), 3.78-3.65 (m, 1H), 3.33 (t, 1H),3.19-3.17 (m, 1H).

Example 21: Synthesis of(S)-2,2-difluoro-4-phenyl-7-((trifluoromethyl)sulfonyl)-2,3-dihydro-1H-inden-1-ol(Compound 487)

(S)-4-bromo-2,2-difluoro-7-((trifluoromethyl)sulfonyl)-2,3-dihydro-1H-inden-1-ol(20 mg, 0.050 mmol) was dissolved in 1,4-dioxane (0.20 mL) and treatedwith phenylboronic acid (9.6 mg, 0.080 mmol), Pd(dppf)Cl₂-DCM adduct (3mg, 0.004 mmol), and potassium fluoride (6.1 mg, 0.10 mmol). The mixturewas heated to 100° C. for 10 hours. After cooling, the reaction mixturewas concentrated in a stream of nitrogen gas then chromatographed onSiO₂ eluting with a gradient of ethyl acetate/hexanes. The desiredmaterial was concentrated to white solid, (17 mg, 86%). LCMS ESI (−) m/z(M−H) 377; ¹H NMR (400 MHz, CDCl₃): δ 8.03 (d, 1H), 7.67 (d, 1H),7.54-7.47 (m, 3H), 7.42-7.39 (m, 2H), 5.47-5.43 (m, 1H), 3.80-3.67 (m,1H), 3.40-3.31 (t, 1H), 3.22-3.21 (m, 1H).

Example 22: Synthesisof(S)-3-(2,2-difluoro-1-hydroxy-7((trifluoromethyl)sulfonyl)-2,3-dihydro-1H-inden-4-yl)-5-fluorobenzonitrile(Compound 486)

(S)-4-bromo-2,2-difluoro-7-((trifluoromethyl)sulfonyl)-2,3-dihydro-1H-inden-1-ol(20 mg, 0.050 mmol) was dissolved in 1,4-dioxane (0.20 mL) and treatedwith 3-cyano-5-fluorophenylboronic acid (10 mg, 0.060 mmol),Pd(dppf)Cl₂-DCM adduct (3 mg, 0.004 mmol), and potassium fluoride (6.1mg, 0.10 mmol). The mixture was heated to 100° C. for 10 hours. Aftercooling, the mixture was concentrated in a stream of nitrogen gas, thendirectly chromatographed on SiO₂ eluting with a gradient of ethylacetate/hexanes. The desired product was recovered as colorless film,(12 mg, 54%). LCMS ESI (−) m/z (M−H) 420; ¹H NMR (400 MHz, CDCl₃): δ8.09 (d, 1H), 7.65 (d, 1H), 7.61-7.60 (m, 2H), 7.40-7.37 (m, 1H),5.49-5.46 (m, 1H), 3.78-3.65 (m, 1H), 3.33-3.25 (t, 1H), 3.20 (m, 1H).

Example 23: Synthesis of(R)-4-((3-chloro-5-fluorophenyl)thio)-7-((difluoromethyl)sulfonyl)-2,3-dihydro-1H-inden-1-ol(Compound 493)

Sodium bicarbonate (18.9 mg, 0.23 mmol) was added all at once to(1R)-7-(difluoromethyl sulfonyl)-4-fluoro-indan-1-ol (20.0 mg, 0.08mmol) and 3-chloro-5-fluoro-benzenethiol (24.4 mg, 0.15 mmol) in1-methyl-2-pyrrolidone (0.5 mL) at room temperature then the reactionvial was sealed with a threaded cap. The reaction mixture was thenwarmed to 90° C. and continued to stir at this temperature untilcomplete as judged by LC-MS (4 h). Cooled to room temperature thenpurified directly on reverse phase silica gel (25+M, 14 CV, 20-100%MeCN/water) affording(1R)-4-(3-chloro-5-fluoro-phenyl)sulfanyl-7-(difluoromethylsulfonyl)indan-1-ol(25.5 mg, 0.062 mmol, 83% yield). LC-MS ESI (−) m/z 453/455 (M+HCO₂ ⁻).¹H-NMR (400 MHz, CDCl₃): δ 7.84 (d, 1H), 7.19-7.17 (m, 1H), 7.08 (s,1H), 7.00-6.97 (m, 2H), 5.71-5.68 (m, 1H), 3.64 (d, 1H), 3.21 (s, 3H),3.12-3.04 (m, 1H), 2.84-2.76 (m, 1H), 2.52-2.43 (m, 1H), 2.27-2.19 (m,1H).

Example 24: Synthesis of(1R)-7-(difluoromethylsulfonyl)-4-(tetrahydropyran-4-ylamino)indan-1-ol(Compound 494)

4-Piperidone (8.4 mg, 0.08 mmol) was added all at once to(1R)-7-(difluoromethyl sulfonyl)-4-fluoro-indan-1-ol (22.0 mg, 0.08mmol) in 1-methyl-2-pyrrolidone (0.5 mL) at room temperature. Thereaction mixture was then stirred at 50° C. for 24 h. Additional4-piperidone (8.4 mg, 0.08 mmol) was added at room temperature and thenwarmed to 90° C. for an additional 4 h. Cooled to room temperature andadded additional 4-piperidone (8.4 mg, 0.08 mmol). Warmed to 90° C. foran additional 4 h. Cooled to room temperature then purified directly onreverse phase silica gel (12+M, 14 CV, 20-100% MeCN/water) affording(1R)-7-(difluoromethylsulfonyl)-4-(tetrahydropyran-4-ylamino)indan-1-ol(15.8 mg, 0.046 mmol, 55% yield). LC-MS ESI (−) m/z 346 (M−H). ¹H-NMR(400 MHz, CDCl₃): δ 7.69 (d, 1H), 6.64 (d, 1H), 6.26 (t, 1H), 5.57-5.56(m, 1H), 4.14 (d, 1H), 4.06-4.01 (m, 2H), 3.72-3.62 (m, 1H), 3.58-3.51(m, 2H), 3.32 (d, 1H), 2.95-2.84 (m, 1H), 2.61-2.55 (m, 1H), 2.47-2.38(m, 1H), 2.28-2.20 (m, 1H), 2.09-2.03 (m, 2H), 1.62-1.52 (m, 2H).

Example 25: Synthesis of4-(2-hydroxyethyl)-7-((trifluoromethyl)sulfonyl)-2,3-dihydro-1H-inden-1-ol(Compound 495)

Step A: Preparation of diethyl2-[7′-(trifluoromethylsulfonyl)spiro[1,3-dioxolane-2,1′-indane]-4′-yl]propanedioate

Tetrahydrofuran (12.0 mL) was added all at once to sodium hydride (735.6mg, 18.39 mmol) at 0° C. under nitrogen followed by the slow addition ofdiethyl malonate (1.86 mL, 12.26 mmol). Stirred for 15 min then asolution of4′-fluoro-7′-(trifluoromethylsulfonyl)spiro[1,3-dioxolane-2,1′-indane](1.0 g, 3.07 mmol) in tetrahydrofuran (3.0 mL) was added by syringe over2 minutes. The reaction mixture was then removed from the cooling bathand stirred at room temperature overnight. Additional sodium hydride(200 mg) was added as well as diethyl malonate (0.5 mL) and stirred anadditional 6 h. Cooled to 0° C., quenched with water, extracted withethyl acetate, washed with brine, dried over sodium sulfate, filteredand concentrated in vacuo. Purified on silica gel (25 g SNAP Ultra,10-100% ethyl acetate/hexanes) affording diethyl2-[7′-(trifluoromethylsulfonyl)spiro[1,3-dioxolane-2,1′-indane]-4′-yl]propanedioate(940.0 mg, 2.01 mmol, 66% yield).

Step B: Preparation of2-[1-oxo-7-(trifluoromethylsulfonyl)indan-4-yl]acetic acid

HCl (4.84 mL, 29.03 mmol) was added to diethyl2-[7′-(trifluoromethylsulfonyl)spiro[1,3-dioxolane-2,1′-indane]-4′-yl]propanedioate(300.0 mg, 0.64 mmol) then warmed to 100° C. for 6 h. Cooled to roomtemperature, extracted with MTBE, washed with water, brine, dried overNa₂SO₄, filtered and concentrated in vacuo affording2-[1-oxo-7-(trifluoromethylsulfonyl)indan-4-yl]acetic acid (200.0 mg,0.62 mmol, 96% yield). Used without further purification.

Step C: Preparation of4-(2-hydroxyethyl)-7-(trifluoromethylsulfonyl)indan

1-ol: Borane dimethylsulfide complex (434.4 μL, 0.87 mmol) was addedslowly to 2-[1-oxo-7-(trifluoromethylsulfonyl)indan-4-yl]acetic acid(70.0 mg, 0.22 mmol) in tetrahydrofuran (1.5 mL) at room temperature andstirred for 2 h. Cooled to 0° C. and quenched with 1 N HCl, extractedwith ethyl acetate, washed with brine, dried over MgSO₄, filtered andconcentrated in vacuo. Purified on silica gel (10 g SNAP Ultra, 14 CV,60-100% ethyl acetate/hexanes) affording4-(2-hydroxyethyl)-7-(trifluoromethylsulfonyl)indan-1-ol (36.0 mg, 0.12mmol, 53% yield). Hexanes was added to the clear oil and then cooled to−78° C. with scratching until a white gum was observed, warmed to roomtemperature and continued scratching until a white powder formed.Hexanes was then removed under a stream of nitrogen to afford Compound495. LC-MS (−) ESI m/z 309 (M−H). ¹H-NMR (400 MHz, CDCl₃): δ 7.83 (d,1H), 7.47 (d, 1H), 5.61 (d, 1H), 3.95-3.92 (m, 1H), 3.27-3.18 (m, 1H),3.10 (s, 1H), 2.99-2.96 (m, 3H), 2.41-2.26 (m, 2H).

Example 26: Synthesis of(S)-2,2-difluoro-4-(2-hydroxyethyl)-7-((trifluoromethyl)sulfonyl)-2,3-dihydro-1H-inden-1-ol(Compound 496)

Step A: Borane dimethylsulfide complex (439.0 μL, 0.88 mmol) was addeddropwise to2-[7′-(trifluoromethylsulfonyl)spiro[1,3-dioxolane-2,1′-indane]-4′-yl]aceticacid (268.0 mg, 0.73 mmol) in tetrahydrofuran (7.0 mL) at 0° C. undernitrogen then slowly warmed to room temperature. Stirred until completeas judged by LC-MS. Quenched carefully with saturated sodiumbicarbonate, extracted with ethyl acetate, washed with brine, dried overMgSO₄, filtered and concentrated in vacuo. Purified on silica gel (10 gSNAP Ultra, 14 CV, 40-100% ethyl acetate/hexanes) affording2-[7′-(trifluoromethylsulfonyl)spiro[1,3-dioxolane-2,1′-indane]-4′-yl]ethanol(95.0 mg, 0.27 mmol, 37% yield).

Step B: HCl (1.0 mL, 1.0 mmol) was added all at once to2-[7′-(trifluoromethylsulfonyl)spiro[1,3-dioxolane-2,1′-indane]-4′-yl]ethanol(95.0 mg, 0.27 mmol) in acetone (4.0 mL) at room temperature thenstirred until complete as judged by LC-MS. Diluted with water, extractedwith ethyl acetate, washed with saturated sodium bicarbonate, washedwith brine, dried over MgSO₄, filtered and concentrated in vacuo. Usedwithout further purification.

Step C: Tert-Butyldimethylsilyl chloride (46.9 mg, 0.31 mmol) was addedall at once to a solution of4-(2-hydroxyethyl)-7-(trifluoromethylsulfonyl)indan-1-one (80.0 mg, 0.26mmol) and imidazole (53.0 mg, 0.78 mmol) in dichloromethane (2.0 mL) atroom temperature then stirred overnight. Diluted with water, extractedwith MTBE, washed with brine, dried over Na₂SO₄, filtered andconcentrated in vacuo. Purified on silica gel (10 g SNAP, 12 CV, 5-60%ethyl acetate/hexanes) affording4-[2-[tert-butyl(dimethyl)silyl]oxyethyl]-7-(trifluoromethylsulfonyl)indan-1-one(89.0 mg, 0.21 mmol, 81% yield).

Step D: Pivalic acid (2.2 mg, 0.02 mmol) was added to a mixture of4-[2-[tert-butyl(dimethyl)silyl]oxyethyl]-7-(trifluoromethylsulfonyl)indan-1-one(89.0 mg, 0.21 mmol) and 3-methoxypropylamine (37.6 mg, 0.42 mmol) incyclohexane (1.5 mL): toluene (1.5 mL) at room temperature then warmedto reflux with the azeotropic removal of water by Dean-Stark trap.Monitored by ¹H-NMR. Cooled to room temperature then concentrated invacuo. Used without further purification.

Step E: Crude4-[2-[tert-butyl(dimethyl)silyl]oxyethyl]-N-(3-methoxypropyl)-7-(trifluoromethylsulfonyl)indan-1-imine(103.0 mg, 0.21 mmol) in acetonitrile (1.5 mL) was added to1-chloromethyl-4-fluoro-1,4-diazoniabicyclo[2.2.2]octanebis(tetrafluoroborate) (184.8 mg, 0.52 mmol) and sodium Sulfate (59.3mg, 0.42 mmol) in acetonitrile (1.5 mL) at 60° C. and stirred for 1 h.Cooled to room temperature then 1 N HCl (3.0 mL) was added and stirredovernight. Extracted with ethyl acetate, washed with brine, dried overMgSO₄, filtered and concentrated in vacuo. Purified on reverse phasesilica gel (12+M, 14 CV, 20-100% acetonitrile/water) affording2,2-difluoro-4-(2-hydroxyethyl)-7-(trifluoromethylsulfonyl)indan-1-one(40.0 mg, 0.12 mmol, 56% yield).

Step F:Chloro{[(1R,2R)-(−)-2-amino-1,2-diphenylethyl](4-toluenesulfonyl)amido}(p-cymene)ruthenium(II)(1.5 mg, 0.002 mmol) was added all at once to an ice cold solution of2,2-difluoro-4-(2-hydroxyethyl)-7-(trifluoromethylsulfonyl)indan-1-one(40.0 mg, 0.12 mmol), triethylamine (32.4 μL, 0.23 mmol) and formic acid(13.2 μL, 0.35 mmol) in dichloromethane (1.0 mL) then sealed with athreaded teflon cap and placed in a 4° C. fridge over the weekend.Purified directly on silica gel (10 g SNAP Ultra, 14 CV, 25-100% ethylacetate/hexanes) affording (1S)-2,2-difluoro-4-(2-hydroxyethyl)-7-(trifluoromethylsulfonyl)indan-1-ol(Compound 496) (28.0 mg, 0.081 mmol, 70% yield) as a clear oil. Swirledwith hexanes to yield a white solid. LC-MS ESI (−) m/z 345 (M−H).

Example 27: Synthesis of3-(2,2-difluoro-1-hydroxy-7-((trifluoromethyl)sulfonyl)-2,3-dihydro-1H-inden-4-yl)propane-1,2-diol(Compound 521)

Step A: Tetrakis(triphenylphosphine)palladium(0) (59.69 mg, 0.0500 mmol)was added all at once to a degassed solution of4′-bromo-7′-(trifluoromethylsulfonyl)spiro[1,3-dioxolane-2,1′-indane](200.0 mg, 0.52 mmol) and allyltributyltin (0.19 mL, 0.62 mmol) in DMF(5.0 mL) under nitrogen then warmed to 90° C. until complete as judgedby LC-MS. Cooled to room temperature, saturated KF (5.0 mL) was addedand stirred for 30 min, extracted with MTBE, washed with brine, driedover MgSO₄, filtered and concentrated in vacuo. Purified on silica gel(10 g SNAP Ultra, 14 CV, 5-50% ethyl acetate/hexanes) affording4′-allyl-7′-(trifluoromethylsulfonyl)spiro[1,3-dioxolane-2,1′-indane](145.0 mg, 0.42 mmol, 81% yield).

Step B: HCl (2.0 mL, 2.0 mmol) was added all at once to a solution of4′-allyl-7′-(trifluoromethylsulfonyl)spiro[1,3-dioxolane-2,1′-indane](145.0 mg, 0.42 mmol) in acetone (5.0 mL) then stirred overnight at roomtemperature. Diluted with water, extracted with MTBE, washed with brine,dried over MgSO₄, filtered and concentrated in vacuo. Used withoutfurther purification.

Step C: Pivalic acid (2.6 mg, 0.02 mmol) was added all at once to4-allyl-7-(trifluoromethylsulfonyl)indan-1-one (76.0 mg, 0.25 mmol) and3-methoxypropylamine (76.4 μL, 0.75 mmol) in cyclohexane (1.5 mL):toluene (1.5 mL) at room temperature then warmed to reflux withazeotropic removal of water via a Dean-Stark trap until complete asjudged by ¹H-NMR. Cooled to room temperature then concentrated in vacuo.Used without further purification.

Step D: 1-chloromethyl-4-fluoro-1,4-diazoniabicyclo[2.2.2]octanebis(tetrafluoroborate) (219.4 mg, 0.62 mmol) was added all at once tocrude4-allyl-N-(3-methoxypropyl)-7-(trifluoromethylsulfonyl)indan-1-imine(93.0 mg, 0.25 mmol) and sodium sulfate (70.4 mg, 0.50 mmol) inacetonitrile (3.0 mL) at 60° C. then stirred for 1 h. Cooled to roomtemperature, 1 N HCl was added (3.0 mL) and stirred for 15 min,extracted with MTBE, washed with brine, dried over MgSO₄, filtered andconcentrated in vacuo. Used without further purification.

Step E: Sodium borohydride (25.0 mg, 0.66 mmol) was added all at once toa solution of4-allyl-2,2-difluoro-7-(trifluoromethylsulfonyl)indan-1-one (75.0 mg,0.22 mmol) and2,2-difluoro-4-[(E)-prop-1-enyl]-7-(trifluoromethylsulfonyl)indan-1-one(75.0 mg, 0.22 mmol) in methanol (2.0 mL) at room temperature andstirred for 30 min. Quenched with 1 N HCl (2.0 mL), diluted with water,extracted with MTBE, washed with brine, dried over Na₂SO₄, filtered andconcentrated in vacuo. Used without further purification.

Step F: Osmium tetroxide, 4 wt. % solution (27.9 μL, 0.004 4 mmol) wasadded all at once to a solution of4-allyl-2,2-difluoro-7-(trifluoromethylsulfonyl)indan-1-ol (75.0 mg,0.22 mmol), 4-methylmorpholine N-oxide (51.3 mg, 0.44 mmol) in acetone(2.0 mL) then stirred over the weekend in a sealed vial. Diluted withwater, extracted with ethyl acetate, washed with brine dried overNa₂SO₄, filtered and concentrated in vacuo. Purified on silica gel (10 gSNAP Ultra, 14 CV, 50-100% ethyl acetate/hexanes) affording Compound 521(17.4 mg, 0.046 mmol, 21% yield). LC-MS ESI (−) m/z 375 (M−H)

Example 28: Synthesis of4-(3-hydroxypropyl)-7-((trifluoromethyl)sulfonyl)-2,3-dihydro-1H-inden-1-ol(Compound 522)

Step A: 3-Ethoxy-3-oxopropylzinc bromide (0.77 mL, 0.39 mmol) was addedto a solution of4′-bromo-7′-(trifluoromethylsulfonyl)spiro[1,3-dioxolane-2,1′-indane](50.0 mg, 0.13 mmol), palladium(II) acetate (2.9 mg, 0.01 mmol) andSPhos (10.6 mg, 0.03 mmol) in tetrahydrofuran (0.5 mL) at roomtemperature under nitrogen in a sealed microwave vial then warmed to 60°C. until complete as judged by LC-MS. Cooled to room temperature,quenched with saturated ammonium chloride, extracted with ethyl acetate,washed with brine, dried over MgSO₄, filtered and concentrated in vacuo.Purified on silica gel (10 g SNAP Ultra, 14 CV, 5-50% ethylacetate/hexanes) affording ethyl3-[7′-(trifluoromethylsulfonyl)spiro[1,3-dioxolane-2,1′-indane]-4′-yl]propanoate(25.0 mg, 0.061 mmol, 47% yield).

Step B: 1 N HCl (1.0 mL, 1.0 mmol) was added all at once to a solutionof ethyl3-[7′-(trifluoromethylsulfonyl)spiro[1,3-dioxolane-2,1′-indane]-4′-yl]propanoate(25.0 mg, 0.06 mmol) in acetone (2.0 mL) then stirred at roomtemperature until complete as judged by LC-MS (30 min). Diluted withbrine, extracted with ethyl acetate, dried over Na₂SO₄, filtered andconcentrated in vacuo. Used without further purification.

Step C: Lithium borohydride solution (0.2 mL, 0.40 mmol) was added to asolution of crude ethyl3-[1-oxo-7-(trifluoromethylsulfonyl)indan-4-yl]propanoate (22.0 mg, 0.06mmol) in tetrahydrofuran (0.50 mL) at room temperature under nitrogenthen stirred until complete as judged by LC-MS. Warmed to 75° C. after 5h at room temperature and held for 3 h. Cooled to room temperature,poured into 1 N HCl, extracted with ethyl acetate, washed with brine,dried over MgSO₄, filtered and concentrated in vacuo. Purified on silicagel (10 g SNAP Ultra, 14 CV, 50%-100% EtOAc/hexanes) affording Compound522 (4.4 mg, 0.014 mmol, 22% yield). LC-MS ESI (−) m/z 323 (M−H).

Example 29: Synthesis of2-[1-oxo-7-(trifluoromethylsulfonyl)indan-4-yl]acetamide (Compound 523)

Step A: Preparation of2-[7′-(trifluoromethylsulfonyl)spiro[1,3-dioxolane-2,1′-indane]-4′-yl]aceticacid

Sodium hydroxide (0.55 mL, 1.66 mmol) added by syringe to diethyl2-[7′-(trifluoromethylsulfonyl)spiro[1,3-dioxolane-2,1′-indane]-4′-yl]propanedioate(515.0 mg, 1.10 mmol) in ethanol 95% (2.2 mL) at room temperature andstirred for 30 min. Warmed to 60° C. for 3 h. Added additional sodiumhydroxide (0.55 mL, 1.66 mmol) and stirred until complete as judged byLC-MS. Acidified to pH 2 with 1 N HCl, diluted with brine, extractedwith ethyl acetate, dried over Na₂SO₄, filtered and concentrated invacuo affording2-[7′-(trifluoromethylsulfonyl)spiro[1,3-dioxolane-2,1′-indane]-4′-yl]aceticacid (368 mg, 1.00 mmol, 91% yield). Used without further purification.

Step B: Preparation of2-[1-oxo-7-(trifluoromethylsulfonyl)indan-4-yl]acetamide

N,N-Diisopropylethylamine (142.7 μL, 0.82 mmol) added all at once to2-[7′-(trifluoromethylsulfonyl)spiro[1,3-dioxolane-2,1′-indane]-4′-yl]aceticacid (100.0 mg, 0.27 mmol), ammonium chloride (73.0 mg, 1.4 mmol) andHATU (156.1 mg, 0.41 mmol) in DMF (2.0 mL) at room temperature thenstirred until complete as judged by LC-MS. Purified directly on reversephase silica gel (12+M, 14 CV, 20-100% acetonitrile/water) affording2-[1-oxo-7-(trifluoromethylsulfonyl)indan-4-yl]acetamide (12.0 mg, 0.037mmol, 14% yield). LC-MS ESI (+) m/z 322 (M+H).

Step C: Preparation of2-[1-hydroxy-7-(trifluoromethylsulfonyl)indan-4-yl]acetamide

Sodium borohydride (4.2 mg, 0.11 mmol) was added all at once to2-[1-oxo-7-(trifluoromethylsulfonyl)indan-4-yl]acetamide (12.0 mg, 0.04mmol) in methanol (1.0 mL) at room temperature and stirred for 20 min.Quenched with 1 N HCl, extracted with ethyl acetate, washed with brine,dried over Na₂SO₄, filtered and concentrated in vacuo. A portion waspurified by preparative TLC (ethyl acetate) affording Compound 523 (1.6mg, 0.005 mmol, 13% yield). LC-MS ESI (−) m/z 322 (M−H).

Example 30: Synthesis of(R)-2-(1-hydroxy-7-((trifluoromethyl)sulfonyl)-2,3-dihydro-1H-inden-4-yl)aceticacid (Compound 524)

Chloro{[(1R,2R)-(−)-2-amino-1,2-diphenylethyl](4-toluenesulfonyl)amido}(p-cymene)ruthenium(II)(2.0 mg, 0.003 mmol) was added all at once to an ice cold solution of2-[1-oxo-7-(trifluoromethylsulfonyl)indan-4-yl]acetic acid (102.0 mg,0.32 mmol), triethylamine (88.2 μL, 0.63 mmol) and formic acid (35.8 μL,0.95 mmol) in dichloromethane (3.0 mL), sealed with a teflon lined capand placed in a 4° C. fridge overnight. Warmed to room temperature thenstirred for an additional 3 days. Concentrated in vacuo then purified onreverse phase silica gel (25+M, 14 CV, 20-100% acetonitrile/water),diluted with ethyl acetate, washed with 1 N HCl to remove triethylamine,washed with brine, dried over Na₂SO₄, filtered and concentrated in vacuoaffording 2-[(1R)-1-hydroxy-7-(trifluoromethylsulfonyl)indan-4-yl]aceticacid (45.0 mg, 0.14 mmol, 44% yield). LC-MS ESI (−) m/z 323 (M−H).

Example 31: Synthesis of(1S,3R)-4-(3,3-difluorocyclobutoxy)-2,2,3-trifluoro-7-(methylsulfonyl)-2,3-dihydro-1H-inden-1-ol(Compound 574) and(1S,3S)-4-(3,3-difluorocyclobutoxy)-2,2,3-trifluoro-7-(methylsulfonyl)-2,3-dihydro-1H-inden-1-ol(Compound 575)

Step A: Preparation of 4,7-difluoro-1H-indene-1,3(2H)-dione: A solutionof 3,6 difluorophthalic anhydride (4.25 g, 23.1 mmol), tert-butyl3-oxobutanoate (4.29 mL, 25.9 mmol) and acetic anhydride (21.0 mL, 221.6mmol) at 25° C. was treated with triethylamine (11.7 mL, 84.3 mmol) andstirred at ambient temperature for 18 h. The reaction mixture was cooledto 0° C. and treated with 10% hydrochloric acid (65 mL, 211 mmol) bydropwise addition. Once the addition was complete, the ice bath wasremoved and the mixture stirred at ambient for 10 minutes. The mixturewas then heated to 75° C. for 10 minutes. During this time gas evolutionwas observed. The suspension slowly broke up to form a clear redmixture. The reaction mixture was poured into 100 mL of water andextracted with 3×50 mL CH₂Cl₂. The combined organics were dried withMgSO₄, filtered, and concentrated to dryness. The product was usedwithout further purification.

Step B: Preparation of 2,2,4,7-tetrafluoro-1H-indene-1,3(2H)-dione: Asolution of the unpurified 4,7-difluoro-1H-indene-1,3(2H)-dione (4.2 g,23.1 mmol) in acetonitrile (100 mL) cooled in a 25° C. water bath wastreated with sodium carbonate (5.38 g, 50.7 mmol). Selectfluor® (17.97g, 50.7 mmol) was added and the reaction mixture was stirred at ambienttemperature for 1 hour. Volatiles were removed under reduced pressureand the residue was poured into 100 mL of 0.1% HCl and extracted with3×50 mL EtOAc. The combined organics were rinsed with 40 mL of brine,dried with MgSO₄, filtered, and concentrated to dryness. The residue waspurified by flash chromatography on silica gel 1:1 hexane/ethyl acetateto give 2,2,4,7-tetrafluoro-1H-indene-1,3(2H)-dione (3.5 g, 70%) as asolid. ¹H NMR (400 MHz, CDCl₃): δ 7.70 (t, 2H).

Step C: Preparation of(S)-2,2,4,7-tetrafluoro-3-hydroxy-2,3-dihydro-1H-inden-1-one: To asolution of 2,2,4,7-tetrafluoro-1H-indene-1,3(2H)-dione (3.48 g, 16.0mmol) in dichloromethane (150 mL) at 0° C. was added formic acid (600μL, 16.0 mmol) and triethylamine (1.55 mL, 11.2 mmol). The resultingmixture was sparged with nitrogen for 5 minutes and thenRuCl(p-cymene)[(S,S)-Ts-DPEN] (203.6 mg, 0.32 mmol) was added. Thereaction vessel was sealed and put into a 4° C. refrigerator to standfor 18 hours. The reaction mixture was poured into 40 mL 1 N HCl. TheCH₂Cl₂ layer was separated and the aqueous layer extracted with ethylacetate (2×50 mL). The combined organics were dried with Na₂SO₄,filtered and concentrated under reduced pressure. The residue waspurified by flash chromatography on silica gel using 25% EtOAc/hexane togive (S)-2,2,4,7-tetrafluoro-3-hydroxy-2,3-dihydro-1H-inden-1-one (2.9g, 83%) as an oil. ¹H NMR (400 MHz, CDCl₃): δ 7.51 (ddd, 1H), 7.29-7.23(m, 1H), 5.44 (dd, 1H), 2.79 (dd, 1H).

Step D: Preparation of(S)-2,2,4-trifluoro-3-hydroxy-7-(methylthio)-2,3-dihydro-1H-inden-1-one:A solution of(S)-2,2,4,7-tetrafluoro-3-hydroxy-2,3-dihydro-1H-inden-1-one (966 mg,4.39 mmol) in acetonitrile (40 mL) at 0° C. was sparged with nitrogenfor 5 minutes and treated with sodium thiomethoxice (353.7 mg, 5.05mmol). The ice bath was removed and the reaction mixture was allowed tostir at ambient temperature for 2 hours. The reaction mixture wasevaporated and the residue partitioned between 40 mL of EtOAc and 40 mLof water. The aqueous layer was further extracted with 2×40 mL of EtOAc.The combined organic extracts were washed with brine, dried over MgSO₄,filtered, and evaporated. The residue was chromatographed on silicausing 10-60% EtOAc/hexane to afford(S)-2,2,4-trifluoro-3-hydroxy-7-(methylthio)-2,3-dihydro-1H-inden-1-one(870 mg, 80%) as a yellow solid. LCMS ESI (+)[M+H]⁺ m/z 249.

Step E: Preparation of(S)-2,2,4-trifluoro-3-hydroxy-7-(methylsulfonyl)-2,3-dihydro-1H-inden-1-one:(S)-2,2,4-trifluoro-3-hydroxy-7-(methylthio)-2,3-dihydro-1H-inden-1-one(400 mg, 1.6 mmol) was dissolved in MeOH (10 mL) and the reaction wastreated dropwise with a solution of Oxone® (2.18 g, 3.55 mmol) dissolvedin water (10 mL). The mixture was stirred at ambient temperature for 14hours. The reaction mixture was filtered, the solids were washed withethyl acetate and the filtrate was concentrated in vacuo. The aqueousfiltrate was extracted 3×30 mL of EtOAc and then the combined organicswere washed with saturated NaCl, dried over Na₂SO₄ and concentrated invacuo to a yellow solid that was used without further purification (467mg). LCMS ESI (+) [M+H]⁺ m/z 281.

Step F: Preparation of(R)-2,2,3,4-tetrafluoro-7-(methylsulfonyl)-2,3-dihydro-1H-inden-1-one:(S)-2,2,4-trifluoro-3-hydroxy-7-(methylsulfonyl)-2,3-dihydro-1H-inden-1-one(450 mg, 1.6 mmol) was dissolved in dichloromethane (16 mL), cooled to0° C. and treated dropwise with diethylaminosulfur trifluoride (0.32 mL,2.4 mmol) and the mixture was stirred at 0° C. for 2 hours, then thewhole homogeneous reaction mixture was placed into the refrigeratorovernight. The reaction was treated with additional diethylaminosulfurtrifluoride (0.32 mL, 2.4 mmol) and stirring continued for 6 h at 0° C.The cold reaction was treated with saturated NaHCO₃ (10 mL) and stirredvigorously for 20 minutes. The mixture was diluted with additionalmethylene chloride and the layers were separated. The aqueous wasre-extracted with methylene chloride and the combined organic layerswere dried over Na₂SO₄ and concentrated in vacuo to a yellow solid. Thecrude material was chromatographed on SiO₂ (Biotage SNAP Ultra) andeluted with a gradient of ethyl acetate/hexane. The desired material wasconcentrated to a pale yellow solid (258 mg). LCMS ESI (+) [M+H]⁺ m/z283.

Step G: Preparation of(R)-2,2,3,4-tetrafluoro-7-(methylsulfonyl)-2,3-dihydrospiro[indene-1,2′-[1,3]dioxolane]:A solution of (3R)-2,2,3,4-tetrafluoro-7-methylsulfonyl-indan-1-one(2.03 g, 7.2 mmol) and 2-bromoethanol (1.53 mL, 21.6 mmol) in DMF (16mL) at 25° C. was treated with potassium carbonate (2.98 g, 21.6 mmol)and stirred at 25° C. for 30 min. The reaction mixture was poured into200 mL of water and extracted with 3×50 mL Et2O. The combined organicswere rinsed with 30 mL of brine, dried with MgSO4, filtered, andconcentrated to dryness. The off-white solid was used without furtherpurification. LCMS ESI (+) [M+H]⁺ m/z 327.

Step H: Preparation of(R)-4-(3,3-difluorocyclobutoxy)-2,2,3-trifluoro-7-(methylsulfonyl)-2,3-dihydrospiro[indene-1,2′-[1,3]dioxolane]:A solution of(3′R)-2′,2′,3′,4′-tetrafluoro-7′-methylsulfonyl-spiro[1,3-dioxolane-2,1′-indane](1.95 g, 5.98 mmol) and 3,3-difluoro-cyclobutanol (770 μL, 7.95 mmol) inacetonitrile (30 mL) at 25° C. was treated with potassium hydroxide(402.4 mg, 7.17 mmol) and stirred at 25° C. for 1 h. Excess acetonitrilewas removed by concentration under reduced pressure. The reactionmixture was poured into 40 mL of water and extracted with 3×40 mL EtOAc.The combined organics were rinsed with 10 mL of brine, dried with MgSO₄,filtered, and concentrated to dryness. Purification was achieved bychromatography on silica using 15-45% EtOAc/hexane to afford a whitesolid (2.2 g, 89%). LCMS ESI (+) [M+H]⁺ m/z 415.

Step I: Preparation of(R)-4-(3,3-difluorocyclobutoxy)-2,2,3-trifluoro-7-(methylsulfonyl)-2,3-dihydro-1H-inden-1-one: A solution of(3′R)-4′-(3,3-difluorocyclobutoxy)-2′,2′,3′-trifluoro-7′-methylsulfonyl-spiro[1,3-dioxolane-2,1′-indane](2.2 g, 5.31 mmol) in dichloromethane (30 mL) at 25° C. was treated withperchloric acid (70% in water, 10 mL) and left to stir for 2 days. Thereaction mixture was carefully quenched by the addition of 100 mL ofsaturated aqueous NaHCO₃ and extracted with 3×50 mL CH₂Cl₂. The combinedorganics were rinsed with 10 mL of brine, dried with MgSO₄, filtered,and concentrated to dryness. Purification was achieved by chromatographyon silica using 25-65% EtOAc/hexane to afford a solid (1.41 g, 72%).LCMS ESI (+) [M+H]⁺ m/z 371.

Step J: Preparation of(1S,3R)-4-(3,3-difluorocyclobutoxy)-2,2,3-trifluoro-7-(methylsulfonyl)-2,3-dihydro-1H-inden-1-ol(Compound 574) and(1S,3S)-4-(3,3-difluorocyclobutoxy)-2,2,3-trifluoro-7-(methylsulfonyl)-2,3-dihydro-1H-inden-1-ol(Compound 575): A solution of(3R)-4-(3,3-difluorocyclobutoxy)-2,2,3-trifluoro-7-methylsulfonyl-indan-1-one(1.41 g, 3.81 mmol) in dichloromethane (40 mL) was cooled to 0° C. andsparged with nitrogen for 5 minutes. During this time formic acid (430μL, 11.42 mmol) and triethylamine (1.06 mL, 7.62 mmol) were sequentiallyadded. Once the sparging was complete, RuCl(p-cymene)[(R,R)-Ts-DPEN](48.5 mg, 0.076 mmol) was added under a continuous stream of nitrogen.The reaction vessel was sealed and put into the refrigerator to reactovernight. Volatiles were removed by concentration under reducedpressure. The residue was purified by chromatography on silica using25-55% EtOAc/hexane. Additional purifications by chromatography onsilica using 20-50% EtOAc/hexane were necessary to isolate material ofsufficient purity. A flash crystallization was preformed from CHCl₃. Thesample was dissolved in a minimum of refluxing CHCl₃ and then cooled to0° C. The collected solid was rinsed with CHCl₃ and dried under highvacuum overnight to afford Compound 574 as a white solid (550 mg, 39%).From the repeated purifications, Compound 575 was isolated as a whitesolid. Data for Compound 574: LCMS ESI (+) [M+H]⁺ m/z 373; ¹H NMR (400MHz, (CD₃)₂CO): δ 8.11 (dd, 1H), 7.33 (d, 1H), 5.87 (dd, 1H), 5.65-5.59(m, 1H), 5.17-5.08 (m, 1H), 3.40-3.26 (m, 2H), 3.27 (s, 3H), 2.98-2.81(m, 2H), 2.80 (t, 1H). Data for Compound 575: LCMS ESI (+) [M+H]⁺ m/z373; ¹H NMR (400 MHz, CDCl₃): δ 8.06 (dd, 1H), 6.87 (d, 1H), 5.92 (dd,1H), 5.78 (td, 1H), 4.86-4.76 (m, 1H), 3.98 (d, 1H), 3.25-3.14 (m, 2H),3.22 (s, 3H), 2.95-2.78 (m, 2H).

Example 32: Synthesis of(1S,3R)-4-(3,3-difluorocyclobutoxy)-2,2,3-trifluoro-7-((fluoromethyl)sulfonyl)-2,3-dihydro-1H-inden-1-ol(Compound 576)

Step A: Preparation of(R)-2,2,3,4-tetrafluoro-7-(methylthio)-2,3-dihydro-1H-inden-1-one: Asolution of(S)-2,2,4-trifluoro-3-hydroxy-7-(methylthio)-2,3-dihydro-1H-inden-1-one(402 mg, 1.62 mmol) in dichloromethane (16.2 mL) at 0° C. was treatedwith diethylaminosulfur trifluoride (390 μL, 2.92 mmol). The ice bathwas removed from the resulting reaction mixture and the reaction mixturewas stirred for 2 hours at room temperature. Volatiles were removed byconcentration under reduced pressure. The residue was suspended in 30 mLof EtOAc, cooled to 0° C., and quenched by the addition of 20 mL ofsaturated aqueous NaHCO₃. The reaction mixture was vigorously stirredfor 30 minutes and then extracted with 3×20 mL EtOAc. The combinedorganics were rinsed with 10 mL of brine, dried with MgSO₄, filtered,and concentrated to dryness. The product was used without furtherpurification. LCMS ESI (+) [M+H]⁺ m/z 251.

Step B: Preparation of(R)-2,2,3,4-tetrafluoro-7-((fluoromethyl)thio)-2,3-dihydro-1H-inden-1-one:A solution of(R)-2,2,3,4-tetrafluoro-7-(methylthio)-2,3-dihydro-1H-inden-1-one (393mg, 1.57 mmol) in acetonitrile (15.7 mL) at 0° C. was treated withSelectfluor® (584.3 mg, 1.65 mmol) and stirred at 0° C. for 2 hours.Volatiles were removed by concentration under reduced pressure. Thereaction mixture was poured into 30 mL of water and extracted with 3×20mL EtOAc. The combined organics were rinsed with 10 mL of brine, driedwith MgSO₄, filtered, and concentrated to dryness. Purification wasachieved by chromatography on silica using 10-30% EtOAc/hexane to afford(R)-2,2,3,4-tetrafluoro-7-((fluoromethyl)thio)-2,3-dihydro-1H-inden-1-one(153 mg, 36%) as a yellow oil. LCMS ESI (+) [M-F]⁺ m/z 249.

Step C: Preparation of(R)-2,2,3,4-tetrafluoro-7-((fluoromethyl)sulfonyl)-2,3-dihydro-1H-inden-1-one:A solution of(R)-2,2,3,4-tetrafluoro-7-((fluoromethyl)thio)-2,3-dihydro-1H-inden-1-one(91.8 mg, 0.34 mmol) in a mixture of methanol (3.4 mL) and water (3.4mL) was treated with Oxone® (252.5 mg, 0.41 mmol). The resultingsuspension was heated to 60° C. overnight. Additional Oxone® (252.5 mg,0.41 mmol) was added and the reaction mixture heated for an additional 6hours. Volatiles were removed by concentration under reduced pressure.The reaction mixture was poured into 100 mL of water and extracted with3×25 mL EtOAc. The combined organics were rinsed with 10 mL of brine,dried with MgSO₄, filtered, and concentrated to dryness. Purificationwas achieved by chromatography on silica using 10-40% EtOAc/hexane toafford(R)-2,2,3,4-tetrafluoro-7-((fluoromethyl)sulfonyl)-2,3-dihydro-1H-inden-1-oneas a white solid (73 mg, 71%). LCMS ESI (+) [M+H]⁺ m/z 301.

Step D: Preparation of(1S,3R)-4-(3,3-difluorocyclobutoxy)-2,2,3-trifluoro-7-((fluoromethyl)sulfonyl)-2,3-dihydro-1H-inden-1-ol(Compound 576): Prepared similarly as described in Example 31, StepsG-J. Purification was achieved by chromatography on silica using 15-30%EtOAc/hexane to afford Compound 576 (29.8 mg, 49%) as a white solid.Retention time HPLC (14 min)=4.63 min; LCMS ESI (+) [M+H]⁺ m/z 391; ¹HNMR (400 MHz, CDCl₃): δ 8.13 (dd, 1H), 6.93 (d, 1H), 5.76 (dd, 1H),5.59-5.53 (m, 1H), 5.47 (dd, 1H), 5.18 (dd, 1H), 4.90-4.80 (m, 1H),3.29-3.16 (m, 2H), 3.16 (d, 1H), 2.97-2.81 (m, 2H).

Example 33: Synthesis of3-(((5R,6S,7S)-5,6-difluoro-7-hydroxy-1-(trifluoromethyl)-6,7-dihydro-5H-cyclopenta[c]pyridin-4-yl)oxy)-5-fluorobenzonitrile(Compound 813)

Step A: Preparation of3-((5-bromo-1-(trifluoromethyl)-5,6-dihydrospiro[cyclopenta[c]pyridine-7,2′-[1,3]dioxolan]-4-yl)oxy)-5-fluorobenzonitrile

A suspension of3-fluoro-5-[1′-(trifluoromethyl)spiro[1,3-dioxolane-2,7′-5,6-dihydrocyclopenta[c]pyridine]-4′-yl]oxy-benzonitrile(1520 mg, 4 mmol), 2,2′-azobisisobutyronitrile (66 mg, 0.4 mmol),magnesium oxide (483 mg, 12 mmol) and N-bromosuccinimide (925 mg, 5.2mmol) in 1,2-dichloroethane (40 mL) was sparged with nitrogen for 3minutes. The vessel was sealed and heated to 80° C. for 3 h. Additional2,2′-azobisisobutyronitrile (66 mg, 0.4 mmol) and N-bromosuccinimide(925 mg, 5.2 mmol) was added to help drive the reaction to completion.Heating was continued for an additional 1.5 h. The reaction mixture wasleft at room temperature overnight. MgO was removed by filtrationthrough the celite and rinsing of the filter cake with CH₂Cl₂. Thefiltrate was treated with 30 mL of saturated aqueous NaHCO₃, stirred for10 minutes, and extracted with 3×30 mL CH₂Cl₂. The combined organicswere rinsed with 10 mL of brine, dried with MgSO₄, filtered, andconcentrated to dryness. Purification was achieved by chromatography onsilica using 5-25% EtOAc/hexane to afford3-((5-bromo-1-(trifluoromethyl)-5,6-dihydrospiro[cyclopenta[c]pyridine-7,2′-[1,3]dioxolan]-4-yl)oxy)-5-fluorobenzonitrileas a yellow foam (931 mg, 51%). LCMS ESI (+) (M+H) m/z 459/461.

Step B: Preparation of3-fluoro-5-((5-hydroxy-1-(trifluoromethyl)-5,6-dihydrospiro[cyclopenta[c]pyridine-7,2′-[1,3]dioxolan]-4-yl)oxy)benzonitrile

A solution of3-[5′-bromo-1′-(trifluoromethyl)spiro[1,3-dioxolane-2,7′-5,6-dihydrocyclopenta[c]pyridine]-4′-yl]oxy-5-fluoro-benzonitrile(931 mg, 2 mmol) in a mixture of 1,2-dimethoxyethane (15 mL) and water(5 mL) at 25° C. was treated with silver(I) carbonate (1.12 g, 4.1 mmol)and stirred at 85° C. for 8 h. An additional portion of silver(I)carbonate (1.12 g, 4.05 mmol) was added and the reaction mixture left toheat at 85° C. overnight. The reaction mixture was diluted with EtOAcand filtered through celite. The filtrate was washed with water andbrine, dried and concentrated. Purification was achieved bychromatography on silica using 20-55% EtOAc/hexane to afford3-fluoro-5-((5-hydroxy-1-(trifluoromethyl)-5,6-dihydrospiro[cyclopenta[c]pyridine-7,2′-[1,3]dioxolan]-4-yl)oxy)benzonitrileas a yellow solid (295 mg, 37%). LCMS ESI (+) (M+H) m/z 397.

Step C: Preparation of3-fluoro-5-((5-oxo-1-(trifluoromethyl)-5,6-dihydrospiro[cyclopenta[c]pyridine-7,2′-[1,3]dioxolan]-4-yl)oxy)benzonitrile

A solution of3-fluoro-5-[5′-hydroxy-1′-(trifluoromethyl)spiro[1,3-dioxolane-2,7′-5,6-dihydrocyclopenta[c]pyridine]-4′-yl]oxy-benzonitrile(296 mg, 0.75 mmol) in dichloromethane (15 mL) at 25° C. was treatedwith Dess-Martin periodinane (396 mg, 0.93 mmol). After 1 h, anadditional 20 mg of Dess-Martin periodinane was added. After stirringfor another 30 minutes, the reaction was quenched by the addition of 6mL of saturated Na₂S₂O₃ solution and 6 mL of saturated NaHCO₃ solution.The resulting biphase was stirred for 10 minutes. The reaction mixturewas poured into 10 mL of water and extracted with 3×20 mL CH₂Cl₂. Thecombined organics were rinsed with 10 mL of brine, dried with MgSO₄,filtered, and concentrated to dryness. The product,3-fluoro-5-((5-oxo-1-(trifluoromethyl)-5,6-dihydrospiro[cyclopenta[c]pyridine-7,2′-[1,3]dioxolan]-4-yl)oxy)benzonitrile(264 mg), was used without further purification. LCMS ESI (+) (M+H) m/z395.

Step D: Preparation of3-fluoro-5-((6-fluoro-5-oxo-1-(trifluoromethyl)-5,6-dihydrospiro[cyclopenta[c]pyridine-7,2′-[1,3]dioxolan]-4-yl)oxy)benzonitrile

A solution of3-fluoro-5-[5′-oxo-1′-(trifluoromethyl)spiro[1,3-dioxolane-2,7′-6H-cyclopenta[c]pyridine]-4′-yl]oxy-benzonitrile(294 mg, 0.75 mmol) and triethylamine (520 μL, 3.7 mmol) indichloromethane (15 mL) at 0° C. was treated withtert-butyldimethylsilyl trifluoromethane (690 μL, 2.98 mmol) and stirredat 0° C. for 30 minutes. The reaction mixture was left to stir for 5 hduring which it was slowly warmed to room temperature. The reactionmixture was poured into 15 mL of saturated NaHCO₃, stirred for 10minutes, and extracted with 3×15 mL CH₂Cl₂. The combined organics wererinsed with 10 mL of brine, dried with MgSO₄, filtered, and concentratedto dryness. The crude residue was dissolved in 4 mL of acetonitrile andtreated with Selectfluor® (264 mg, 0.75 mmol). The reaction mixture wasstirred for 3 h at room temperature. Volatiles were removed byconcentration under reduced pressure. The mixture was poured into 30 mLof water and extracted with 3×15 mL EtOAc. The combined organics wererinsed with 10 mL of brine, dried with MgSO₄, filtered, and concentratedto dryness. Purification was achieved by chromatography on silica using5-30% EtOAc/hexane to afford3-fluoro-5-((6-fluoro-5-oxo-1-(trifluoromethyl)-5,6-dihydrospiro[cyclopenta[c]pyridine-7,2′-[1,3]dioxolan]-4-yl)oxy)benzonitrileas a white solid (206 mg, 67%). ¹H NMR (400 MHz, CDCl₃): δ 8.51 (s, 1H),7.32-7.28 (m, 1H), 7.21-7.18 (m, 1H), 7.13-7.08 (m, 1H), 5.20 (d, 1H),4.51-4.31 (m, 4H).

Step E: Preparation of3-fluoro-5-(((5R,6R)-6-fluoro-5-hydroxy-1-(trifluoromethyl)-5,6-dihydrospiro[cyclopenta[c]pyridine-7,2′-[1,3]dioxolan]-4-yl)oxy)benzonitrile

A solution of3-fluoro-5-[6′-fluoro-5′-oxo-1′-(trifluoromethyl)spiro[1,3-dioxolane-2,7′-6H-cyclopenta[c]pyridine]-4′-yl]oxy-benzonitrile(206 mg, 0.5 mmol) in dichloromethane (10 mL) was cooled to 0° C. andsparged with nitrogen for 5 minutes. During this time, formic acid (57μL, 1.50 mmol) and triethylamine (104 μL, 0.75 mmol) were sequentiallyadded. Once sparging was complete, RuCl(p-cymene)[(S,S)-Ts-DPEN] (6.4mg, 0.01 mmol) was added under a continuous stream of nitrogen. Thereaction vessel was sealed and put into the refrigerator to reactovernight. Volatiles were removed by concentration under reducedpressure. Purification was achieved by chromatography on silica using20-45% EtOAc/hexane to afford3-fluoro-5-(((5R,6R)-6-fluoro-5-hydroxy-1-(trifluoromethyl)-5,6-dihydrospiro[cyclopenta[c]pyridine-7,2′-[1,3]dioxolan]-4-yl)oxy)benzonitrileas a clear solid (200 mg, 97%). LCMS ESI (+) (M+H) m/z 415.

Step F: Preparation of(5S,6R)-4-(3-cyano-5-fluorophenoxy)-6-fluoro-1-(trifluoromethyl)-5,6-dihydrospiro[cyclopenta[c]pyridine-7,2′-[1,3]dioxolan]-5-yl4-nitrobenzoate

A solution of3-fluoro-5-[(5′R,6′R)-6′-fluoro-5′-hydroxy-1′-(trifluoromethyl)spiro[1,3-dioxolane-2,7′-5,6-dihydrocyclopenta[c]pyridine]-4′-yl]oxy-benzonitrile(200 mg, 0.48 mmol), polymer supported triphenylphosphine (˜2.06 mmol/g,938 mg, 1.93 mmol), and 4-nitrobenzoic acid (323 mg, 1.93 mmol) intetrahydrofuran (9.7 mL) at 25° C. was treated with diisopropylazodicarboxylate (371 μL, 1.88 mmol) and stirred for 2 h. The reactionmixture was filtered and the filter cake rinsed with EtOAc. The filtratewas concentrated and purified by chromatography on silica using 10-35%EtOAc/hexane to afford(5S,6R)-4-(3-cyano-5-fluorophenoxy)-6-fluoro-1-(trifluoromethyl)-5,6-dihydrospiro[cyclopenta[c]pyridine-7,2′-[1,3]dioxolan]-5-yl4-nitrobenzoate as a white solid (221 mg, 81%). LCMS ESI (+) (M+H) m/z564.

Step G: Preparation of3-fluoro-5-(((5S,6R)-6-fluoro-5-hydroxy-1-(trifluoromethyl)-5,6-dihydrospiro[cyclopenta[c]pyridine-7,2′-[1,3]dioxolan]-4-yl)oxy)benzonitrile

A solution of[(5'S,6′R)-4′-(3-cyano-5-fluoro-phenoxy)-6′-fluoro-1′-(trifluoromethyl)spiro[1,3-dioxolane-2,7′-5,6-dihydrocyclopenta[c]pyridine]-5′-yl]4-nitrobenzoate (238 mg, 0.42 mmol) in tetrahydrofuran (8 mL) at 25° C.was treated with a solution of freshly prepared lithium hydroxidehydrate (19.5 mg, 0.46 mmol) in water (2.4 mL) and stirred at 25° C. for30 minutes. An additional portion of lithium hydroxide hydrate (10 mg,0.24 mmol) in water (1.2 mL) was added and the reaction mixture stirredfor 30 minutes. 2.0 mL of saturated NH₄Cl was added to the reactionmixture and volatiles were removed under reduced pressure. The reactionmixture was poured into 10 mL of saturated NaHCO₃ and extracted with3×15 mL EtOAc. The combined organics were rinsed with 10 mL of brine,dried with MgSO₄, filtered, and concentrated to dryness. Purificationwas achieved by chromatography on silica using 5-10%EtOAc/dichloromethane to afford3-fluoro-5-(((5S,6R)-6-fluoro-5-hydroxy-1-(trifluoromethyl)-5,6-dihydrospiro[cyclopenta[c]pyridine-7,2′-[1,3]dioxolan]-4-yl)oxy)benzonitrileas a white solid (137 mg, 78%). LCMS ESI (+) (M+H) m/z 415.

Step H: Preparation of3-(((5R,6S)-5,6-difluoro-1-(trifluoromethyl)-5,6-dihydrospiro[cyclopenta[c]pyridine-7,2′-[1,3]dioxolan]-4-yl)oxy)-5-fluorobenzonitrile

A solution of3-fluoro-5-[(5'S,6′R)-6′-fluoro-5′-hydroxy-1′-(trifluoromethyl)spiro[1,3-dioxolane-2,7′-5,6-dihydrocyclopenta[c]pyridine]-4′-yl]oxy-benzonitrile(137 mg, 0.33 mmol) in dichloromethane (6.6 mL) at 25° C. was treatedwith diethylaminosulfur trifluoride (87.4 L, 0.66 mmol). The reactionmixture was allowed to stir for 30 minutes. The reaction mixture wasquenched by the careful addition of 2 mL of aqueous saturated NaHCO₃.The resulting mixture vigorously stirred for 30 minutes. The reactionmixture was poured into 20 mL of water and extracted with 3×10 mLCH₂Cl₂. The combined organics were rinsed with 10 mL of brine, driedwith MgSO₄, filtered, and concentrated to dryness. Purification wasachieved by chromatography on silica using 10-30% EtOAc/hexane to afford3-(((5R,6S)-5,6-difluoro-1-(trifluoromethyl)-5,6-dihydrospiro[cyclopenta[c]pyridine-7,2′-[1,3]dioxolan]-4-yl)oxy)-5-fluorobenzonitrileas a white solid (92.7 mg, 67%). LCMS ESI (+) (M+H) m/z 417.

Step I: Preparation of3-(((5R,6S,7S)-5,6-difluoro-7-hydroxy-1-(trifluoromethyl)-6,7-dihydro-5H-cyclopenta[c]pyridin-4-yl)oxy)-5-fluorobenzonitrile(Compound 813)

A solution of3-[(5′R,6'S)-5′,6′-difluoro-1′-(trifluoromethyl)spiro[1,3-dioxolane-2,7′-5,6-dihydrocyclopenta[c]pyridine]-4′-yl]oxy-5-fluoro-benzonitrile(93 mg, 0.22 mmol) in dichloromethane (5 mL) at 0 C was treated with 70%aqueous perchloric acid (1.0 mL) and stirred at 44 C for 3 h. Thereaction mixture was cooled to 0° C., carefully quenched with a mixtureof 10 mL of saturated NaHCO₃/10 mL of water and extracted with 3×15 mLCH₂Cl₂. The combined organics were rinsed with 10 mL of brine, driedwith MgSO₄, filtered, and concentrated to dryness. The intermediateketone product was used immediately without further purification bydissolving in 4 mL of MeOH, cooling to 0° C., and treating with sodiumborohydride (8.4 mg, 0.22 mmol). The reaction stirred for 15 min and wasthen quenched by the addition of 1 mL of saturated NH₄Cl. Volatiles wereremoved by concentration under reduced pressure. The reaction mixturewas poured into 20 mL of water and extracted with 3×10 mL EtOAc. Thecombined organics were rinsed with 10 mL of brine, dried with MgSO₄,filtered, and concentrated to dryness. Purification was achieved bychromatography on silica using 10-45% EtOAc/hexane to afford3-(((5R,6S,7S)-5,6-difluoro-7-hydroxy-1-(trifluoromethyl)-6,7-dihydro-5H-cyclopenta[c]pyridin-4-yl)oxy)-5-fluorobenzonitrileas a white solid (60 mg, 72%). Retention time HPLC (14 min)=4.18minutes; LCMS ESI (+) (M+H) m/z 375; ¹H NMR (400 MHz, CDCl₃): δ 8.42 (s,1H), 7.28 (ddd, 1H), 7.20-7.18 (m, 1H), 7.09 (dt, 1H), 5.92 (dt, 1H),5.53-5.46 (m, 1H), 5.19 (ddt, 1H), 2.66 (ddd, 1H).

Example 34: Synthesis of5-(((5R,6S,7S)-5,6-difluoro-7-hydroxy-1-(trifluoromethyl)-6,7-dihydro-5H-cyclopenta[c]pyridin-4-yl)oxy)nicotinonitrile(Compound 825)

Step A: Preparation of5-(((5R,6S,7S)-5,6-difluoro-7-hydroxy-1-(trifluoromethyl)-6,7-dihydro-5H-cyclopenta[c]pyridin-4-yl)oxy)nicotinonitrile(Compound 825)

A solution of tetrahydrofuran (2.0 mL) and Water (4.0 mL) was spargedwith nitrogen for 3 minutes. Sparging was ceased and to the solutionwere sequentially added under continuous nitrogen stream(5R,6S,7S)-4-[(5-bromo-3-pyridyl)oxy]-5,6-difluoro-1-(trifluoromethyl)-6,7-dihydro-5H-cyclopenta[c]pyridin-7-ol(50 mg, 0.12 mmol),[2-(2-aminophenyl)phenyl]-methylsulfonyloxy-palladium;ditert-butyl-[2-(2,4,6-triisopropylphenyl)phenyl]phosphane (19 mg, 0.024mmol) and zinc cyanide (20 mg, 0.17 mmol). The vessel was sealed andheated to 40° C. for 3 h. An additional portion of[2-(2-aminophenyl)phenyl]-methyl sulfonyloxy-palladium;ditert-butyl-[2-(2,4,6-triisopropylphenyl)phenyl]phosphane (19 mg, 0.024mmol) and zinc cyanide (20 mg, 0.17 mmol) were added and the reactionmixture was heated to 65° C. for 18 h. The reaction mixture was pouredinto 10 mL of saturated NaHCO₃ and extracted with 3×10 mL EtOAc. Thecombined organics were rinsed with 10 mL of brine, dried with MgSO₄,filtered, and concentrated to dryness. Purification was achieved bychromatography on silica using 20-60% EtOAc/hexane to afford5-(((5R,6S,7S)-5,6-difluoro-7-hydroxy-1-(trifluoromethyl)-6,7-dihydro-5H-cyclopenta[c]pyridin-4-yl)oxy)nicotinonitrileas a white solid (34.3 mg, 80%). Retention time HPLC (14 min)=2.55minutes; LCMS ESI (+) (M+H) m/z 358; ¹H NMR (400 MHz, CDCl₃): δ 8.79(dd, 1H), 8.71 (d, 1H), 8.41 (s, 1H), 7.67 (dd, 1H), 5.94 (dt, 1H),5.53-5.46 (m, 1H), 5.21 (ddt, 1H), 2.73 (ddd, 1H).

Example 35: Synthesis of(5R,6S,7S)-4-(3,3-difluorocyclobutoxy)-5,6-difluoro-1-(trifluoromethyl)-6,7-dihydro-5H-cyclopenta[c]pyridin-7-ol(Compound 828)

Step A: Preparation of4-(3,3-difluorocyclobutoxy)-1-(trifluoromethyl)-5,6-dihydrospiro[cyclopenta[c]pyridine-7,2′-[1,3]dioxolane]

A solution of1′-(trifluoromethyl)spiro[1,3-dioxolane-2,7′-5,6-dihydrocyclopenta[c]pyridine]-4′-ol(1.9 g, 7.27 mmol), (3,3-difluorocyclobutyl) 4-methylbenzenesulfonate(2.86 g, 10.9 mmol), potassium iodide (1.81 g, 10.9 mmol) and potassiumcarbonate (2.0 g, 14.6 mmol) in acetonitrile (20 mL) was stirred at 100°C. overnight. The reaction mixture was concentrated to dryness, dilutedwith ethyl acetate (100 mL), washed with water (100 mL) and brine (20mL). The organic phase was collected, dried over anhydrous Na₂SO₄,filtered and the filtrate was concentrated. Purification was achieved bychromatography on silica using 10-20% EtOAc/petroleum ether to afford4′-(3,3-difluorocyclobutoxy)-1′-(trifluoromethyl)spiro[1,3-dioxolane-2,7′-5,6-dihydrocyclopenta[c]pyridine]as a white solid (1.00 g, 2.85 mmol, 39%). LCMS ESI (+) (M+H) m/z 352.(The Mitsunobu reaction can also be used to derivatize the1′-(trifluoromethyl)spiro[1,3-dioxolane-2,7′-5,6-dihydrocyclopenta[c]pyridine]-4′-ol.)

Step B: Preparation of5-bromo-4-(3,3-difluorocyclobutoxy)-1-(trifluoromethyl)-5,6-dihydrospiro[cyclopenta[c]pyridine-7,2′-[1,3]dioxolane]

A solution of4′-(3,3-difluorocyclobutoxy)-1′-(trifluoromethyl)spiro[1,3-dioxolane-2,7′-5,6-dihydrocyclopenta[c]pyridine](1.00 g, 2.85 mmol), 2,2′-azobisisobutyronitrile (94 mg, 0.57 mmol),NaHCO₃ (420 mg, 5.0 mmol) and 1-bromopyrrolidine-2,5-dione (1.27 g, 7.12mmol) in 1,2-dichloroethane (20 mL) was sparged with nitrogen for 3minutes. The vessel was sealed and heated to 80° C. for 1 h. Thereaction mixture was poured into 30 mL of saturated aqueous Na₂SO₃ andextracted with 3×30 mL CH₂Cl₂. The combined organics were rinsed with 20mL of brine, dried with MgSO₄, filtered, and concentrated to dryness.Purification was achieved by chromatography on silica using 5-25%EtOAc/hexane to afford5′-bromo-4′-(3,3-difluorocyclobutoxy)-1′-(trifluoromethyl)spiro[1,3-dioxolane-2,7′-5,6-dihydrocyclopenta[c]pyridine]as an off-white solid (650 mg, 1.5 mmol, 53%). ¹H NMR (400 MHz, CDCl₃):δ 8.11 (s, 1H), 5.36-5.31 (m, 1H), 4.98-4.88 (m, 1H), 4.36-4.21 (m, 2H),4.16-4.07 (m, 2H), 3.26-3.13 (m, 2H), 2.96-2.71 (m, 4H).

Step C: Preparation of4-(3,3-difluorocyclobutoxy)-1-(trifluoromethyl)-5,6-dihydrospiro[cyclopenta[c]pyridine-7,2′-[1,3]dioxolan]-5-ol

A solution of5′-bromo-4′-(3,3-difluorocyclobutoxy)-1′-(trifluoromethyl)spiro[1,3-dioxolane-2,7′-5,6-dihydrocyclopenta[c]pyridine](650 mg, 1.5 mmol) in a mixture of 1,2-dimethoxyethane (15 mL) and water(5 mL) was treated with silver carbonate (417 mg, 1.5 mmol) and stirredat 85° C. overnight. The mixture was diluted with EtOAc and filteredthrough celite. The filtrate was concentrated to remove thedimethoxyethane. The residue was re-suspended in 60 mL of 1:1 EtOAc/H₂Oand extracted with 3×15 mL EtOAc. The combined organics were rinsed with10 mL of brine, dried with MgSO₄, filtered, and concentrated to dryness.Purification was achieved by chromatography on silica using 10-30%EtOAc/hexane to afford4′-(3,3-difluorocyclobutoxy)-1′-(trifluoromethyl)spiro[1,3-dioxolane-2,7′-5,6-dihydrocyclopenta[c]pyridine]-5′-olas a solid (280 mg, 50%). ¹H NMR (400 MHz, CDCl₃): δ 8.11 (s, 1H),5.39-5.32 (m, 1H), 4.97-4.87 (m, 1H), 4.33-4.20 (m, 2H), 4.17-4.04 (m,2H), 3.27-3.13 (m, 2H), 2.96-2.75 (m, 2H), 2.63 (dd, 1H), 2.53 (d, 1H),2.28 (dd, 1H).

Step D: Preparation of4-(3,3-difluorocyclobutoxy)-1-(trifluoromethyl)spiro[cyclopenta[c]pyridine-7,2′-[1,3]dioxolan]-5(6H)-one

A solution of4′-(3,3-difluorocyclobutoxy)-1′-(trifluoromethyl)spiro[1,3-dioxolane-2,7′-5,6-dihydrocyclopenta[c]pyridine]-5′-ol(280 mg, 0.76 mmol) in dichloromethane (10 mL) at 25° C. was treatedwith Dess-Martin periodinane (500 mg, 1.18 mmol). After 2 h, thereaction was quenched by the addition of 10 mL of saturated Na₂S₂O₃solution and 10 mL of saturated NaHCO₃ solution. The resulting biphasestirred for 10 minutes. The reaction mixture was poured into 20 mL ofwater and extracted with 3×20 mL CH₂Cl₂. The combined organics wererinsed with 10 mL of brine, dried with MgSO₄, filtered, and concentratedto dryness to afford4′-(3,3-difluorocyclobutoxy)-1′-(trifluoromethyl)spiro[1,3-dioxolane-2,7′-6H-cyclopenta[c]pyridine]-5′-oneas a solid (270 mg, 97%) that was used without further purification.LCMS ESI (+) (M+H) m/z 366.

Step E: Preparation of4-(3,3-difluorocyclobutoxy)-6-fluoro-1-(trifluoromethyl)spiro[cyclopenta[c]pyridine-7,2′-[1,3]dioxolan]-5(6H)-one

A solution of4′-(3,3-difluorocyclobutoxy)-1′-(trifluoromethyl)spiro[1,3-dioxolane-2,7′-6H-cyclopenta[c]pyridine]-5′-one(270 mg, 0.74 mmol) and triethylamine (410 μL, 3 mmol) indichloromethane (10 mL) at 0° C. was treated withtert-butyl-dimethyl-(trifluoromethylsulfonyl)silane (480 μL, 2.2 mmol)and stirred at 0° C. for 30 minutes. The reaction mixture was left tostir overnight at room temperature. The reaction mixture was poured into10 mL of saturated NaHCO₃, stirred for 10 minutes, and extracted with3×15 mL CH₂Cl₂. The combined organics were rinsed with 10 mL of brine,dried with MgSO₄, filtered, and concentrated to dryness. The unpurifiedresidue was dissolved in acetonitrile (10 mL) and treated withSelectfluor® (322 mg, 0.92 mmol). The reaction stirred for 6 h at roomtemperature. Volatiles were removed by concentration under reducedpressure. The reaction mixture was poured into 30 mL of water andextracted with 3×15 mL EtOAc. The combined organics were rinsed with 10mL of brine, dried with MgSO₄, filtered, and concentrated to dryness.Purification was achieved by chromatography on silica using 5-20%EtOAc/hexane to afford4′-(3,3-difluorocyclobutoxy)-6′-fluoro-1′-(trifluoromethyl)spiro[1,3-dioxolane-2,7′-6H-cyclopenta[c]pyridine]-5′-oneas a white solid (180 mg, 56%). ¹H NMR (400 MHz, CDCl₃): δ 8.35 (s, 1H),5.13 (d, 1H), 5.01-4.91 (m, 1H), 4.47-4.38 (m, 1H), 4.38-4.26 (m, 3H),3.30-3.14 (m, 2H), 3.04-2.86 (m, 2H).

Step F: Preparation of(5R,6R)-4-(3,3-difluorocyclobutoxy)-6-fluoro-1-(trifluoromethyl)-5,6-dihydrospiro[cyclopenta[c]pyridine-7,2′-[1,3]dioxolan]-5-ol

A solution of4′-(3,3-difluorocyclobutoxy)-6′-fluoro-1′-(trifluoromethyl)spiro[1,3-dioxolane-2,7′-6H-cyclopenta[c]pyridine]-5′-one(180 mg, 0.47 mmol) in dichloromethane (10 mL) was cooled to 0° C. andsparged with nitrogen for 5 minutes. During this time formic acid (53.1μL, 1.4 mmol) and triethylamine (98.2 μL, 0.70 mmol) were sequentiallyadded. Once sparging was complete, RuCl(p-cymene)[(S,S)-Ts-DPEN] (15 mg,0.023 mmol) was added under a continuous stream of nitrogen. Thereaction vessel was sealed and put into the refrigerator to reactovernight. Volatiles were removed by concentration under reducedpressure. Purification was achieved by chromatography on silica using10-30% EtOAc/hexane to afford(5′R,6′R)-4′-(3,3-difluorocyclobutoxy)-6′-fluoro-1′-(trifluoromethyl)spiro[1,3-dioxolane-2,7′-5,6-dihydrocyclopenta[c]pyridine]-5′-olas a clear solid (152 mg, 84%). LCMS ESI (+) (M+H) m/z 386.

Step G: Preparation of(5S,6R)-4-(3,3-difluorocyclobutoxy)-6-fluoro-1-(trifluoromethyl)-5,6-dihydrospiro[cyclopenta[c]pyridine-7,2′-[1,3]dioxolan]-5-yl4-nitrobenzoate

A solution of(5′R,6′R)-4′-(3,3-difluorocyclobutoxy)-6′-fluoro-1′-(trifluoromethyl)spiro[1,3-dioxolane-2,7′-5,6-dihydrocyclopenta[c]pyridine]-5′-ol(124 mg, 0.32 mmol), polymer supported triphenylphosphine (˜2.06 mmol/g,627 mg, 1.29 mmol), and 4-nitrobenzoic acid (216 mg, 1.29 mmol) intetrahydrofuran (3.3 mL) was treated with diisopropyl azodicarboxylate(248 μL, 1.26 mmol) and stirred at 25° C. for 2 h. The reaction mixturewas filtered and the filter cake rinsed with 30 mL EtOAc. The filtratewas concentrated and purified by chromatography on silica using 10-25%EtOAc/hexane to afford(5S,6R)-4-(3,3-difluorocyclobutoxy)-6-fluoro-1-(trifluoromethyl)-5,6-dihydrospiro[cyclopenta[c]pyridine-7,2′-[1,3]dioxolan]-5-yl4-nitrobenzoate as a white solid (160 mg, 93%). LCMS ESI (+) (M+H) m/z535.

Step H: Preparation of(5S,6R)-4-(3,3-difluorocyclobutoxy)-6-fluoro-1-(trifluoromethyl)-5,6-dihydrospiro[cyclopenta[c]pyridine-7,2′-[1,3]dioxolan]-5-ol

A solution of[(5'S,6′R)-4′-(3,3-difluorocyclobutoxy)-6′-fluoro-1′-(trifluoromethyl)spiro[1,3-dioxolane-2,7′-5,6-dihydrocyclopenta[c]pyridine]-5′-yl]4-nitrobenzoate (194 mg, 0.36 mmol) in tetrahydrofuran (7 mL) at 25° C.was treated with a solution of freshly prepared hydroxylithium hydrate(16.8 mg, 0.4 mmol) in water (2.1 mL) and stirred at 25° C. for 30minutes. An additional portion of hydroxylithium hydrate (8.4 mg, 0.20mmol) in water (1.0 mL) was added and the reaction mixture stirred foranother 30 minutes. Saturated NH₄Cl (0.5 mL) was added and volatileswere removed under reduced pressure. The reaction mixture was pouredinto 10 mL of saturated NaHCO₃ and extracted with 3×15 mL EtOAc. Thecombined organics were rinsed with 10 mL of brine, dried with MgSO₄,filtered, and concentrated to dryness. Purification was achieved bychromatography on silica using 10-35% EtOAc/hexane to afford(5S,6R)-4-(3,3-difluorocyclobutoxy)-6-fluoro-1-(trifluoromethyl)-5,6-dihydrospiro[cyclopenta[c]pyridine-7,2′-[1,3]dioxolan]-5-olas a white solid (74 mg, 53%). LCMS ESI (+) (M+H) m/z 386.

Step I: Preparation of(5R,6S)-4-(3,3-difluorocyclobutoxy)-5,6-difluoro-1-(trifluoromethyl)-5,6-dihydrospiro[cyclopenta[c]pyridine-7,2′-[1,3]dioxolane]

A solution of(5'S,6′R)-4′-(3,3-difluorocyclobutoxy)-6′-fluoro-1′-(trifluoromethyl)spiro[1,3-dioxolane-2,7′-5,6-dihydrocyclopenta[c]pyridine]-5′-ol(74 mg, 0.19 mmol) in dichloromethane (3.8 mL) at 25° C. was treatedwith diethylaminosulfur trifluoride (51 μL, 0.38 mmol). The reactionmixture was allowed to stir for 30 minutes. The reaction mixture wasquenched by the careful addition of 1 mL of aqueous saturated NaHCO₃.The resulting mixture stirred for 30 minutes, poured into 20 mL of waterand extracted with 3×15 mL CH₂Cl₂. The combined organics were rinsedwith 10 mL of brine, dried with MgSO₄, filtered, and concentrated todryness. Purification was achieved by chromatography on silica using5-25% EtOAc/hexane to afford(5R,6S)-4-(3,3-difluorocyclobutoxy)-5,6-difluoro-1-(trifluoromethyl)-5,6-dihydrospiro[cyclopenta[c]pyridine-7,2′-[1,3]dioxolane]as a white solid (72 mg, 97%). LCMS ESI (+) (M+H) m/z 388.

Step J: Preparation of(5R,6S,7S)-4-(3,3-difluorocyclobutoxy)-5,6-difluoro-1-(trifluoromethyl)-6,7-dihydro-5H-cyclopenta[c]pyridin-7-ol(Compound 828)

A solution of(5′R,6'S)-4′-(3,3-difluorocyclobutoxy)-5′,6′-difluoro-1′-(trifluoromethyl)spiro[1,3-dioxolane-2,7′-5,6-dihydrocyclopenta[c]pyridine](72 mg, 0.19 mmol) in dichloromethane (4.0 mL) at 0° C. was treated with70% aqueous perchloric acid (800 μL). The reaction mixture was heated to44° C. for 5 h. The reaction mixture was cooled to 0° C., carefullyquenched with a mixture of 10 mL of saturated NaHCO₃/10 mL of water andextracted with 3×15 mL CH₂Cl₂. The combined organics were rinsed with 10mL of brine, dried with MgSO₄, filtered, and concentrated to dryness.The product was used immediately without further purification bydissolving in 2 mL of MeOH, cooling to 0° C., and treating with sodiumborohydride (7.0 mg, 0.19 mmol). The reaction stirred for 15 min and wasthen quenched by the addition of 1 mL of saturated NH₄Cl. Volatiles wereremoved by concentration under reduced pressure. The reaction mixturewas poured into 20 mL of water and extracted with 3×15 mL EtOAc. Thecombined organics were rinsed with 10 mL of brine, dried with MgSO₄,filtered, and concentrated to dryness. Purification was achieved bychromatography on silica using 10-40% EtOAc/hexane to afford(5R,6S,7S)-4-(3,3-difluorocyclobutoxy)-5,6-difluoro-1-(trifluoromethyl)-6,7-dihydro-5H-cyclopenta[c]pyridin-7-olas a white solid (53 mg, 83%). Retention time HPLC (14 min)=3.37minutes; LCMS ESI (+) (M+H) m/z 346; ¹H NMR (400 MHz, CDCl₃): δ 8.24 (s,1H), 5.95 (ddd, 1H), 5.47-5.41 (m, 1H), 5.07 (ddt, 1H), 4.97-4.88 (m,1H), 3.30-3.15 (m, 2H), 2.99-2.79 (m, 2H), 2.54-2.49 (m, 1H).

Example 36: HIF-2α Scintillation Proximity Assay (SPA)

The total assay volume was about 100 μL in the following configuration:2 μL compound in 100% DMSO, 88 μL buffer with protein and probe and 10μL of SPA beads. The compound was diluted in a master plate consistingof a 10-point dose response with a 3-fold compound dilution from 100 μMto 5 nM. Assays were run on a 96-well plate in which one column,designated as the high signal control, contained DMSO with no compoundand another column, designated as the low signal control, contained noprotein. Prior to plating out of compound, a buffer solution, consistingof 25 mM TRIS pH 7.5 (Sigma), 150 mM NaCl (Sigma), 15% Glycerol (Sigma),0.15% BSA (Sigma), 0.001% Tween-20 (Sigma), 150 nMN-(3-Chlorophenyl-4,6-t₂)-4-nitrobenzo[c][1,2,5]oxadiazol-5-amine(Compound 183) and 100 nM HIF-2α HIS TAG-PASB Domain, was made andallowed to equilibrate for 30 minutes. Compounds that were to be testedwere then plated in to a 96-well white clear bottom Isoplate-96 SPAplate (Perkin Elmer). To the compounds was added 88 μL of the buffersolution, then the plate covered with a plastic cover and aluminum foil,placed onto a shaker and equilibrated for 1 hour. After equilibration,10 μL of a 2 mg/mL solution of YSi Cu His tagged SPA beads (PerkinElmer) were then added to each well of the plate, covered andequilibrated for another 2 hours. The plates were then removed from theshaker, placed into a 1450 LSC and luminescence counter MicroBeta Trilux(Perkin Elmer) to measure the extent of probe displacement. The percentinhibition was determined and IC₅₀ values were calculated using theDotmatics system based on the following equation: % inhibition=[(highcontrol−sample)/(high control−low control)]×100.

Example 37: VEGF ELISA Assay

About 7500 786-O cells in 180 μL of growth medium were seeded into eachwell of a 96-well, white, clear bottom plate (07-200-566, FisherScientific) on day one in the following layout:

Four hours later, serial dilutions of 10× compound stocks were made ingrowth medium from 500× DMSO stocks, and 20 μL of those 10× stocks wereadded to each well to make final concentrations as follows (μM): 20,6.67, 2.22, 0.74, 0.25, 0.082, 0.027, 0.009, 0.003, 0.001, and 0. Eachconcentration was plated in duplicate. About 20 hours later, medium wasremoved by suction and each well was supplied with 180 μL of growthmedium. About 20 □l freshly-made 10× compound stocks were added to eachwell. About 24 hours later, cell culture medium was removed and the VEGFconcentration determined using an ELISA kit purchased from R&D systems,following the manufacturer's suggested method. The EC₅₀ was calculatedby GraphPad Prism using the dose-response-inhibition (four parameter)equation. The cell-seeded plate was then subjected to CellTiter-Gloluminescence cell viability assay (Promega) by adding 50 μL of CelltiterGlo reagent into each well and shaking the plate for 8 minutes at 550rpm (Thermomixer R, Eppendorf) then the luminescence signal immediatelyread in a plate reader (3 second delay, 0.5 second/well integrationtime, Synergy 2 multi Detection Microplate reader).

Example 38: Luciferase Assay

786-O-Hif-Luc single clone cells were obtained by infecting 786-O cells(ATCC® CRL-1932™) with commercial lentivirus that delivers a luciferasegene driven by multiple HIF responsive elements (Cignal Lenti HIFReporter (luc): CLS-007L, Qiagen) at Multiplicity of Infection (MOI) of25 for 24 hours. The cells were replenished with fresh medium(Dulbecco's Modified Eagle's Medium (DMEM, D5796, Sigma) supplementedwith 10% FBS (F6178, Sigma), 100 units penicillin and 100 μgstreptomycin/mL (P4333, Sigma)) for another 24 hours. A pool of infectedcells were then selected against 2 μg/mL of puromycin (P8833, Sigma) for10 days followed by limited dilution to select single clones. The cloneswere tested for their response to HIF2 inhibitors and the ones thatshowed the biggest dynamic range (786-O-Hif-Luc) were expanded and usedfor the luciferase assay. For the luciferase assay, about 7500786-O-Hif-Luc cells in 90 μL growth medium were seeded into each well ofa 96-well white opaque plate (08-771-26, Fisher scientific) a day beforetreatment with the layout:

On treatment day, serial dilutions of 10× compound stocks were made ingrowth medium from 500×DMSO stocks, and 10 μL of the 10× stocks wereadded to each well to make final concentrations as follows (μM): 20,6.67, 2.22, 0.74, 0.25, 0.08, 0.027, 0.009, 0.003, 0.001, and 0. Eachconcentration was tested in triplicate. After about 24 hours, luciferaseactivity was determined using ONE-Glo Luciferase Assay Reagent (E6110,Promega) following the manufacturer's recommended procedure. EC₅₀ werecalculated using Dotmatics software.

Table 2 shows biological activities of selected compounds in Luciferase,VEGF ELISA and Scintillation Proximity assays. Compound numberscorrespond to the numbers and structures provided in Table 1 andExamples 1-35.

TABLE 2 Less than 50 nM to 250 nM to Greater than 50 nM (++++) 249 nM(+++) 1000 nM (++) 1000 nM (+) Scintillation 1, 2, 6, 7a, 8, 9, 17, 18,21, 33, 38, 3, 5, 16, 22, 24, 4, 7b, 12, 13, 14, Proximity 11, 15, 25,26, 29, 39, 41, 50, 54, 74, 27, 31, 40, 42, 43, 19, 20, 23, 28, 35,Assay 30, 32, 34, 52, 55, 75, 90, 93, 94, 99, 46, 48, 53, 70, 71, 36,37, 44, 45, 47, IC₅₀ (nM) 56, 57, 58, 59, 60, 100, 102, 107, 72, 76, 78,81, 91, 49, 51, 66, 68, 69, 61, 62, 63, 64, 65, 116, 117, 118, 103, 104,106, 73, 77, 79, 82, 83, 67, 80, 92, 98, 119, 128, 136, 115, 120, 131,84, 85, 86, 87, 88, 101, 111, 112, 141, 145, 146, 132, 133, 134, 89, 95,96, 97, 123, 124, 140, 147, 148, 153, 135, 137, 152, 105, 108, 109, 143,144, 155, 156, 162, 165, 157, 172, 178, 110, 113, 114, 158, 159, 160,187, 192, 195, 182, 184, 190, 121, 122, 125, 161, 163, 166, 203, 204,214, 193, 197, 202, 126, 127, 129, 167, 168, 179, 224, 237, 241, 207,209, 212, 130, 138, 139, 185, 186, 188, 242, 252, 254, 213, 218, 220,142, 149, 150, 191, 194, 196, 260, 265, 267, 243, 244, 249, 151, 154,164, 198, 200, 201, 270, 274, 275, 258, 261, 264, 169, 170, 171, 206,215, 221, 276, 277, 285, 269, 271, 281, 173, 174, 175, 223, 225, 227,295, 302, 308, 287, 293, 307, 176, 177, 180, 228, 229, 230, 312, 324,325, 339, 346, 369, 181, 189, 199, 231, 232, 233, 333, 334, 336, 374,411, 422, 205, 208, 210, 234, 235, 236, 353, 358, 361, 423, 432, 433,211, 216, 217, 240, 245, 247, 370, 378, 381, 434, 471, 476, 219, 222,226, 251, 256, 263, 383, 387, 388, 481, 482, 487, 238, 239, 246, 266,273, 286, 399, 405, 409, 496, 505, 506, 248, 250, 253, 289, 290, 292,412, 414, 421, 511, 513, 522, 255, 257, 259, 303, 304, 305, 424, 426,427, 523, 531, 542, 262, 268, 272, 306, 309, 310, 437, 442, 444, 575,580, 591, 278, 279, 280, 314, 315, 316, 462, 468, 477, 595, 601, 603,282, 283, 284, 317, 319, 321, 480, 488, 517, 612, 622, 637, 288, 291,294, 327, 328, 329, 545, 547, 551, 644, 648, 678, 296, 297, 298, 331,338, 340, 554, 563, 569, 703, 707, 711, 299, 300, 301, 341, 342, 344,584, 586, 598, 735, 748, 749, 311, 313, 318, 347, 348, 349, 604, 609,614, 750, 754, 762, 320, 322, 323, 355, 356, 357, 616, 620, 660, 765,770, 775, 326, 330, 332, 359, 360, 364, 663, 669, 672, 781, 812, 816,335, 337, 343, 365, 368, 371, 676, 681, 682, 827 345, 350, 351, 372,375, 376, 686, 696, 697, 352, 354, 362, 379, 386, 389, 700, 701, 706,363, 366, 367, 392, 397, 398, 714, 717, 718, 373, 377, 380, 401, 403,430, 730, 734, 737, 382, 384, 385, 431, 435, 436, 747, 751, 755, 390,391, 393, 440, 443, 446, 759, 768, 769, 394, 395, 396, 451, 458, 465,771, 782, 784, 400, 402, 404, 466, 467, 472, 788, 795, 808, 406, 407,408, 473, 478, 483, 815, 818 410, 413, 415, 485, 486, 489, 416, 417,418, 491, 507, 509, 419, 420, 425, 532, 549, 550, 428, 429, 438, 557,571, 572, 439, 441, 445, 573, 574, 576, 447, 448, 449, 577, 578, 579,450, 452, 453, 581, 582, 585, 454, 455, 456, 587, 593, 594, 457, 459,460, 602, 605, 607, 461, 463, 464, 608, 610, 617, 469, 470, 474, 623,626, 654, 475, 484, 490, 655, 656, 657, 492, 493, 494, 658, 659, 661,495, 497, 498, 675, 677, 699, 499, 500, 501, 704, 709, 710, 502, 503,504, 712, 713, 715, 508, 510, 512, 716, 727, 728, 514, 515, 516, 731,736, 739, 518, 519, 520, 740, 741, 742, 521, 524, 525, 743, 744, 746,526, 527, 528, 753, 756, 760, 529, 530, 533, 761, 766, 774, 534, 535,536, 778, 783, 786, 537, 538, 539, 787, 789, 790, 540, 541, 543, 794,796, 811, 544, 546, 548, 813, 814, 825, 552, 553, 555, 828, 831 556,558, 559, 560, 561, 562, 564, 565, 566, 567, 568, 570, 583, 588, 589,590, 592, 596, 597, 599, 600, 606, 611, 613, 615, 618, 619, 621, 624,625, 627, 628, 629, 630, 631, 632, 633, 634, 635, 636, 638, 639, 640,641, 642, 643, 645, 646, 647, 649, 650, 651, 652, 653, 662, 664, 665,666, 667, 668, 670, 671, 673, 674, 679, 680, 683, 684, 685, 687, 688,689, 690, 691, 692, 693, 694, 695, 698, 702, 705, 708, 719, 720, 721,722, 723, 724, 725, 726, 729, 732, 733, 738, 745, 752, 757, 758, 763,764, 767, 772, 773, 776, 777, 779, 780, 785, 791, 792, 793, 797, 807,809, 810, 817, 819, 820, 821, 822, 823, 824, 826, 829, 830, 833 Mean 8,9, 11, 15, 25, 2, 17, 67, 155, 1, 34, 41, 74, 78, 98, 133, 179, 274 VEGF55, 60, 63, 64, 166, 188, 191, 80, 99, 102, 124, ELISA 158, 159, 161,225, 231, 234, 132, 165, 203, EC₅₀ (nM) 163, 167, 185, 240, 245, 252,267, 353, 387, 186, 196, 228, 254, 256, 303, 424, 473, 495, 230, 233,235, 304, 310, 325, 577, 734 236, 251, 273, 342, 347, 349, 289, 292,305, 355, 357, 360, 306, 309, 316, 365, 372, 398, 317, 364, 368, 399,421, 431, 375, 389, 403, 467, 507, 574, 446, 451, 458, 576, 578, 605,465, 478, 489, 610, 620, 659, 571, 572, 579, 706, 710, 713, 617, 654,655, 736, 740, 743, 656, 657, 658, 760, 761, 783, 709, 712, 742, 784,825 744, 813, 828 Mean 8, 9, 11, 15, 25, 1, 2, 6, 17, 26, 27, 3, 5, 7a,34, 38, 16, 18, 20, 21, 22, Luciferase 55, 63, 64, 65, 56, 57, 58, 59,60, 39, 41, 42, 43, 52, 31, 32, 33, 35, 40, EC₅₀ (nM) 158, 159, 160, 61,62, 67, 155, 54, 74, 80, 81, 90, 46, 50, 53, 75, 85, 161, 163, 166, 162,165, 187, 92, 93, 94, 99, 91, 98, 100, 103, 167, 185, 186, 188, 191,200, 101, 107, 112, 104, 110, 111, 196, 215, 221, 206, 223, 224, 115,124, 144, 114, 116, 117, 225, 227, 228, 237, 241, 245, 145, 146, 156,118, 119, 120, 229, 230, 231, 251, 252, 254, 168, 192, 194, 128, 131,132, 232, 233, 234, 260, 267, 270, 198, 203, 207, 134, 135, 136, 235,236, 240, 274, 276, 277, 213, 242, 243, 140, 143, 147, 247, 256, 266,285, 286, 290, 263, 265, 271, 148, 151, 152, 273, 289, 292, 302, 310,314, 275, 287, 293, 157, 172, 181, 303, 304, 305, 315, 334, 336, 294,295, 308, 182, 190, 195, 306, 309, 316, 342, 348, 355, 312, 324, 325,201, 202, 209, 317, 338, 347, 357, 360, 365, 339, 353, 359, 212, 214,218, 349, 364, 368, 369, 372, 381, 361, 370, 377, 220, 222, 226, 371,375, 380, 383, 387, 388, 378, 390, 411, 244, 249, 250, 386, 389, 399,392, 398, 401, 414, 416, 422, 253, 255, 258, 403, 430, 431, 405, 409,421, 426, 432, 434, 259, 261, 264, 435, 446, 451, 423, 424, 436, 437,440, 443, 268, 269, 272, 458, 465, 467, 473, 477, 480, 444, 462, 466,279, 291, 296, 478, 489, 571, 486, 488, 507, 468, 471, 472, 300, 301,307, 572, 576, 579, 523, 547, 549, 482, 483, 505, 313, 352, 356, 582,584, 602, 573, 574, 578, 517, 531, 532, 358, 363, 374, 605, 617, 654,593, 594, 598, 550, 557, 563, 376, 379, 385, 655, 656, 657, 604, 610,620, 569, 577, 585, 394, 397, 400, 658, 661, 675, 623, 626, 659, 591,595, 608, 412, 427, 428, 704, 709, 710, 676, 677, 681, 616, 622, 644,433, 439, 441, 712, 716, 727, 686, 696, 697, 660, 663, 669, 455, 456,464, 728, 742, 743, 699, 706, 713, 678, 682, 707, 470, 474, 476, 744,746, 783, 715, 717, 730, 711, 714, 718, 481, 485, 487, 786, 787, 789,731, 734, 736, 747, 748, 750, 491, 492, 495, 790, 794, 813, 740, 760,761, 753, 756, 759, 496, 506, 509, 828, 831 781, 782, 784, 762, 766,768, 511, 513, 516, 795, 814, 818, 769, 771, 774, 534, 538, 542, 825775, 788, 796 545, 551, 554, 558, 565, 570, 575, 580, 581, 586, 601,603, 607, 609, 612, 614, 618, 619, 637, 639, 665, 670, 672, 683, 692,700, 701, 703, 721, 722, 723, 724, 725, 726, 735, 737, 739, 741, 749,751, 752, 754, 755, 764, 765, 767, 770, 778, 779, 780, 808, 811, 812,833

Example 39: Antitumor Activity of Compound 163 in Combination withα-PD-1 in Mouse Melanoma B16-F10 Syngeneic Tumor Model

B16-F10 mouse melanoma cells (1×10⁵ cells) in 50 μL of PBS and Matrigel(1:1 in volume) were inoculated subcutaneously in the right flank ofeach C57BL/6 mouse at 6-7 weeks of age for tumor development. When thexenografts reached about 100 mm³ in size, the tumor bearing mice werestratified into four groups (n=8) and treatment was started with vehicle(BID), and Compound 163 (30 mg/kg, BID) in combination with α-PD-1 (10mg/kg, BIW) or IgG control (10 mg/kg, BIW) as indicated in FIG. 1, fortwo weeks. Tumor sizes were measured in two dimensions using a caliperand the volume was expressed in mm³ using the formula V=0.5×a×b², wherea and b were the long and short diameters of the tumor, respectively.All data were displayed as Mean and the standard error of the mean(SEM). FIG. 1 illustrates the observed synergistic effect of thedescribed combination treatment. Table 3 gives the mean tumor size ofeach treatment group after 14 days of treatment.

TABLE 3 Treatment Vehicle + Vehicle + Compound 163 + Compound 163 +groups IgG α-PD-1 IgG α-PD-1 Tumor 3055.05 ± 1958.2 ± 2133.89 ± 306.441018.86 ± 109.23 size 416.62 362.48 (mm³) Mean ± SEM

Example 40: Assay for Testing Inhibition of HIF-2α Dimerization

This example describes an example assay for testing inhibition of HIF-2αdimerization using co-immunoprecipitation.

Cell Culture and Compound Treatment:

786-O cells were cultured in Dulbecco's Modified Eagle's Medium (DMEM,D5796, Sigma) supplemented with 10% FBS (F6178, Sigma), 100 unitspenicillin, and 100 μg/mL streptomycin (P4333, Sigma). About 5×10⁵ cellswere plated into 6-well cell culture plates (Corning Cat#3506) in 2 mLof media and were placed in a 37° C. cell culture incubator withatmospheric levels of oxygen and 5% CO₂. As the cultures reachedconfluence, 2 μl of Compound 163 or 226 diluted in DMSO at 1000× offinal concentration was added.

Co-Immunoprecipitation Analysis of HIF and ARNT:

Cells in a 6-well plate were washed with 1 ml of DPBS and lysed in 1 mlof cell lysis buffer (Tris-HCl 20 mM, pH 7.5, Triton X100 1%, NaCl 150mM, Glycerol 5%, EDTA 1 mM, DTT 1 mM and 1 tablet per 10 mls of RocheProteinase Inhibitor Tablet Complete). The cell lysate was transferredto a 1.5 mL Eppendorf tube, and was incubated on ice for 15 to 20 min.The debris was spun down in an Eppendorf microfuge tube at 15,000 RPMfor 15 min at 4° C., and the supernatant was transferred into a newtube. 1 μg of mouse mAb against human ARNT (Santa Cruz, sc-55526) or 1μg of antibody against HIF-2α (Abcam Ab199) and 50 μl of Protein AGbeads (Santa Cruse, 50% slurry in lysis buffer) were added. The tubeswere rotated at 4° C. for 2 to 16 hours. The beads were spun down andwashed three times with 1 ml cold lysis buffer and resuspended in 30 μlof SDS sample buffer (10% Glycerol, 60 mM Tri-HCl pH6.8, 2% SodiumDodecyl Sulfate, 0.01% Bromophenol Blue and 1.25% β-mercaptoethanol).The tubes were boiled for 3 min. The beads were spun down, andsupernatant was loaded on a denaturing acrylamide gel (Bio-Rad precastedgel) for electrophoresis. The proteins in the gels were then transferredto 0.2 μM PVDF membrane (Bio-Rad) using Bio-Rad Trans-Blot Turbotransfer system.

For Western Blotting, the membranes were incubated in TBST (10 mMTris-HCl pH 7.4, 150 mM Sodium Chloride and 0.1% Tween 20) containing 5%dry milk for 1 hour. The membranes were incubated with antibody toHIF-2α (Abcam Ab199) at 1:500 dilutions in the blocking bufferovernight. The membrane was washed three times in TBST then incubated inTBST with 5% dry milk with HPR-conjugated goat anti rabbit (Bio-Rad172-1019) at 1:5000 dilution for ARNT detection. The Western blotsignals were developed using Thermo Super-Western Pico Lumino/EnhancerSolution following the protocol provided by the manufacturer.

Disruption of HIF-2α and ARNT Dimerization:

Total protein was extracted from 786-O cells treated with the indicatedconcentration of Compound 163 and Compound 226. HIF-2α protein complexwas immunoprecipitated with antibody to HIF-2α. The immunoprecipitantswere blotted with antibody against ARNT. ARNT protein coprecipitatedwith HIF-2α was reduced in a lysate of cells treated with Compound 163in a dose-dependent manner (FIG. 2). Compound 226 reduced ARNT proteinin complex with HIF-2α only at the highest concentration (10 μM).

Example 41: Antitumor Activity of Compound 163 in Combination withα-CTLA-4 in Mouse Colon CT26 Syngeneic Tumor Model

CT26.WT mouse carcinoma cells (5×10⁵ cells) in 50 μL of PBS and Matrigel(1:1 in volume) were inoculated subcutaneously in the right flank ofeach BALB/c mouse at 6-7 weeks of age for tumor development. When thexenografts reached about 90 mm³ in size, the tumor bearing mice werestratified into four groups (n=8) and treatment was started with vehicle(BID), and Compound 163 (30 mg/kg, BID) in combination with α-CTLA-4 (10mg/kg, BIW) or IgG control (10 mg/kg, BIW) as indicated in FIG. 3, for20 days. Tumor sizes were measured in two dimensions using a caliper andthe volume was expressed in mm³ using the formula V=0.5×a×b², where aand b were the long and short diameters of the tumor, respectively. Alldata were displayed as Mean and the standard error of the mean (SEM).FIG. 3 illustrates the observed synergistic effect of the describedcombination treatment. Table 4 gives the mean tumor size of eachtreatment group after 20 days of treatment.

TABLE 4 Compound Compound Treatment Vehicle + Vehicle + 163 + 163 +groups IgG α-CTLA-4 IgG α-CTLA-4 Tumor 1801 ± 190 830 ± 356 1612 ± 305403 ± 108 size (mm³) Mean ± SEM

While preferred embodiments of the present disclosure have been shownand described herein, it will be obvious to those skilled in the artthat such embodiments are provided by way of example only. Numerousvariations, changes, and substitutions will now occur to those skilledin the art without departing from the disclosure. It should beunderstood that various alternatives to the embodiments of thedisclosure described herein may be employed in practicing thedisclosure. It is intended that the appended claims define the scope ofthe disclosure and that methods and structures within the scope of theseclaims and their equivalents be covered thereby.

What is claimed is:
 1. A method of treating a proliferative disorder ina subject in need thereof, comprising administering to said subject aHIF-2α inhibitor in combination with an immunotherapeutic agent.
 2. Themethod of claim 1, wherein the HIF-2α inhibitor and theimmunotherapeutic agent yield a synergistic effect in treating theproliferative disorder.
 3. The method of claim 1, wherein theproliferative disorder is a cancer condition.
 4. The method of claim 3,wherein the cancer condition is selected from the group consisting ofnon-small-cell lung carcinoma, melanoma, renal cell carcinoma,colorectal cancer, castration-resistant prostate cancer, hepatocellularcarcinoma, squamous cell carcinoma of the head and neck, carcinomas ofthe esophagus, ovary, gastrointestinal tract and breast, and ahematologic malignancy.
 5. The method of claim 3, wherein the cancercondition is renal cell carcinoma.
 6. The method of claim 3, wherein theHIF-2α inhibitor and the immunotherapeutic agent together are effectivein one or more of inhibiting proliferation of cancer cells, inhibitingmetastasis of cancer cells, killing cancer cells, modulating an immuneresponse, and reducing severity or incidence of symptoms associated withthe presence of cancer cells.
 7. The method of claim 1, wherein theHIF-2α inhibitor is administered simultaneously with theimmunotherapeutic agent.
 8. The method of claim 1, wherein the HIF-2αinhibitor is administered before exposure to the immunotherapeuticagent.
 9. The method of claim 1, wherein the HIF-2α inhibitor isadministered after exposure to the immunotherapeutic agent.
 10. Themethod of claim 1, wherein the HIF-2α inhibitor or the immunotherapeuticagent are administered parenterally, orally, intraperitoneally,intravenously, intraarterially, transdermally, intramuscularly,liposomally, via local delivery by catheter or stent, subcutaneously,intraadiposally, or intrathecally.
 11. The method of claim 1, whereinthe HIF-2α inhibitor inhibits one or more biological effects selectedfrom the group consisting of heterodimerization of HIF-2α to HIF1β,HIF-2α target gene expression, VEGF gene expression, and VEGF proteinsecretion.
 12. The method of claim 11, wherein the HIF-2α inhibitorinhibits heterodimerization of HIF-2α to HIF1β but notheterodimerization of HIF1α to HIF1β.
 13. The method of claim 1, whereinthe HIF-2α inhibitor binds the PAS-B domain cavity of HIF2α.
 14. Themethod of claim 1, wherein the HIF-2α inhibitor is a compound of FormulaI:

or a pharmaceutically acceptable salt or prodrug thereof, wherein: X isCR⁵ or N; Y is CR⁶ or N; Z is —O—, —S—, —C(HR⁷)—, —N(R⁸)— or absent; R¹is alkyl, heteroalkyl, cycloalkyl, heterocycloalkyl, aryl, heteroaryl,acyl or cyano; R² is nitro, carboxaldehyde, carboxyl, ester, amido,cyano, halo, sulfonyl, alkyl, alkenyl, alkynyl or heteroalkyl; R³ ishydrogen, halo, cyano, alkyl, heteroalkyl, alkenyl, alkynyl, amino,carboxaldehyde, carboxylic acid, oxime, ester, amido or acyl; or R² andR³ taken together form a cyclic moiety; R⁴ is nitro, halo, cyano, alkyl,cycloalkyl, heteroaryl, carboxyl, sulfinyl, sulfonamidyl, sulfonyl orsulfoximinyl; and R⁵, R⁶, R⁷ and R⁸ are independently hydrogen, halo,hydroxy, cyano, alkyl or alkoxy.
 15. The method of claim 14, wherein R²is halo, cyano or alkyl.
 16. The method of claim 14 or 15, wherein R³ ishalo, cyano or alkyl.
 17. The method of any one of claims 14 to 16,wherein R³ is —(CH₂)_(n)OH and n is 1, 2 or
 3. 18. The method of claim17, wherein n is
 1. 19. The method of claim 14, wherein: R² is halo,cyano or alkyl; R³ is halo, cyano or alkyl; and R⁴ is cyano,fluoroalkyl, sulfinyl, sulfonamidyl, sulfonyl or sulfoximinyl.
 20. Themethod of claim 1, wherein the HIF-2α inhibitor is a compound of FormulaI-C:

or a pharmaceutically acceptable salt or prodrug thereof, wherein: X isCR⁵ or N; Y is CR⁶ or N; Z is —O—, —S—, —C(HR⁷)—, —N(R⁸)— or absent; R¹is alkyl, heteroalkyl, cycloalkyl, heterocycloalkyl, aryl, heteroaryl,acyl or cyano; R⁴ is nitro, halo, cyano, alkyl, cycloalkyl, heteroaryl,carboxyl, sulfinyl, sulfonamidyl, sulfonyl or sulfoximinyl; R⁵, R⁶, R⁷and R⁸ are independently hydrogen, halo, hydroxy, cyano, alkyl oralkoxy; R¹¹ is hydrogen, halo, hydroxy, alkoxy or amino; R¹² ishydrogen, alkyl, alkenyl or alkynyl; or R¹¹ and R¹² in combination formoxo or oxime; each of R¹³ is independently selected from the groupconsisting of hydrogen, fluoro, chloro, hydroxy, alkyl and heteroalkyl;or two R¹³s and the carbon atom(s) to which they are attached form a 3-to 8-membered cycloalkyl or heterocycloalkyl moiety; and n is 0, 1, 2, 3or
 4. 21. The method of claim 20, wherein R¹² is hydrogen.
 22. Themethod of claim 20, wherein: R⁴ is cyano, fluoroalkyl, sulfinyl,sulfonamidyl, sulfonyl or sulfoximinyl; R¹¹ is hydroxy or amino; and R¹²is hydrogen.
 23. The method of any one of claims 20 to 22, wherein R¹³is fluoro.
 24. The method of claim 20, wherein the HIF-2α inhibitor is acompound of Formula I-H, I-I, I-J or I-K:

or a pharmaceutically acceptable salt or prodrug thereof.
 25. The methodof any one of claims 20 to 24, wherein R¹¹ is hydroxy or amino.
 26. Themethod of claim 25, wherein R¹¹ is hydroxy.
 27. The method of any one ofclaims 14 to 26, wherein R¹ is phenyl, monocyclic heteroaryl or bicyclicheteroaryl.
 28. The method of claim 27, wherein R¹ is phenyl or pyridyl.29. The method of any one of claims 14 to 26, wherein R¹ is cycloalkylor heterocycloalkyl.
 30. The method of any one of claims 14 to 29,wherein R¹ is substituted with at least one substituent selected fromthe group consisting of halo, C₁-C₄ alkyl, C₁-C₄ alkoxy and cyano. 31.The method of any one of claims 14 to 30, wherein R⁴ is cyano,fluoroalkyl, sulfinyl, sulfonamidyl, sulfonyl or sulfoximinyl.
 32. Themethod of any one of claims 14 to 30, wherein R⁴ is fluoroalkyl oralkylsulfonyl.
 33. The method of any one of claims 14 to 32, wherein Zis —O—.
 34. The method of claim 20, wherein: R⁴ is fluoroalkyl; n is 0,1, 2 or 3; Z is —O—; R¹¹ is hydroxy; and R¹² is hydrogen.
 35. The methodof claim 20, wherein: R⁴ is sulfonyl; n is 0, 1, 2 or 3; Z is —O—; R¹¹is hydroxy; and R¹² is hydrogen.
 36. The method of claim 34 or 35,wherein R¹ is phenyl, pyridyl, cycloalkyl or heterocycloalkyl.
 37. Themethod of any one of claims 14 to 36, wherein X is N and Y is CR⁶. 38.The method of any one of claims 14 to 36, wherein X is CR⁵ and Y is N.39. The method of any one of claims 14 to 36, wherein X is N and Y is N.40. The method of any one of claims 14 to 36, wherein X is CR⁵ and Y isCR⁶.
 41. The method of claim 1, wherein the HIF-2α inhibitor is acompound selected from Table
 1. 42. The method of claim 1, wherein theHIF-2α inhibitor is:

or a pharmaceutically acceptable salt thereof.
 43. The method of claim1, wherein the HIF-2α inhibitor is selected from the group consistingof:

or a pharmaceutically acceptable salt thereof.
 44. The method of any oneof the preceding claims, wherein the immunotherapeutic agent induces orenhances an immune response.
 45. The method of any one of the precedingclaims, wherein the immunotherapeutic agent is a PD-1 inhibitor.
 46. Themethod of claim 45, wherein the PD-1 inhibitor is nivolumab orpembrolizumab.
 47. The method of any one of claims 1 to 44, wherein theimmunotherapeutic agent is a CTLA-4 inhibitor.
 48. The method of claim47, wherein the CTLA-4 inhibitor is tremelimumab or ipilimumab.
 49. Themethod of any one of the preceding claims, wherein the immunotherapeuticagent comprises a small molecule or antibody.
 50. The method of claim49, wherein the immunotherapeutic agent comprises an antibody.
 51. Themethod of any one of the preceding claims, further comprising (a)measuring a tumor size or an amount of one or more markers for thepresence of the proliferative disorder before and after administeringthe HIF-2α inhibitor in combination with the immunotherapeutic agent,and (b) adjusting dosage or discontinuing use of the HIF-2α inhibitorand/or the immunotherapeutic agent based on results of step (a).